Tau imaging probe

ABSTRACT

An object of the present invention is to provide a probe for imaging a β-sheet structure protein which can be used for the diagnosis of conformational diseases, particularly disease (tauopathy) having a cardinal symptom such as intracerebral accumulation of tau protein, for example, Alzheimer&#39;s disease. Another object of the present invention is to provide a compound which is highly specific to tau and can image tau with satisfactory sensitivity, and also has high brain transition, low or non-recognized bone-seeking properties and low or non-recognized toxicity. 
     According to the present invention, the above problems are solved by providing a compound of a formula I (wherein A, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R a  and R b  are as defined in the present description) or a pharmaceutically acceptable salt or solvate thereof.

TECHNICAL FIELD

The present invention relates to a probe for imaging a β-sheet structureprotein which can be used for the diagnosis of conformational diseases,particularly disease (tauopathy) having a cardinal symptom such asintracerebral accumulation of tau protein, for example, Alzheimer'sdisease.

BACKGROUND ART

In Alzheimer's disease, it is known that the accumulation of senileplaque containing amyloid beta-protein (hereinafter referredcollectively to as Aβ) as a main component and of neurofibrillarytangles containing hyperphosphorylated tau protein (hereinafter referredcollectively to as tau) as a main component proceeds to the degree thatit cannot be treated, when the people around the patient or thephysician notice the specific clinical symptoms of the disease. In otherwords, if the current diagnosis of Alzheimer's disease is compared tothat of cancer, it is detected only when it has reached the end stage.

Recently, it has been revealed that even in the case of the extremelyearly stage of very mild Alzheimer's disease that corresponds to mildcognitive impairment (MCI), which is considered as partly precursorstate of Alzheimer's disease, autopsy samples show the accumulation ofmany Aβ and tau, and the state is pathologically almost Alzheimer'sdisease. Therefore, in Alzheimer's disease, the histopathology ismanifested far before the symptom of memory loss appears. In otherwords, there is a quite large difference between the histopathology andclinical picture of Alzheimer's disease (so-called difference betweenpathological Alzheimer's disease and clinical Alzheimer's disease).

As illustrated in FIG. 1, the accumulation of Aβ is considered to start10 or more years earlier than that of tau in the brain of Alzheimer'sdisease. As is apparent from FIG. 1, since the tracing of Aβ wasconsidered most appropriate in order to diagnose Alzheimer's disease inthe extremely early stage or before its' development, almost all PETprobes for the diagnosis of Alzheimer's disease were so-called probesfor amyloid imaging to trace Aβ from the 20th to 21st century. At first,[¹¹C] labeled probes were mainly used, but afterwards, the developmentof [¹⁸F] labeled probes that have a long half-life and are easily usedin the clinical setting has been attempted. FIG. 2 illustrates theexamples of probes for amyloid imaging that have been developed untilnow.

In the beginning of 2002, images showing administration of PET probesfor amyloid imaging to Alzheimer's disease patients were introduced forthe first time in the world (refer to Non-Patent Document 1). The teamof Barrio et al., UCLA got the honor of this, and the probes used were[¹⁸F] FDDNP. Afterwards, however, [¹¹C] PIB developed by GeneralElectric, University of Pittsburgh, which has probably now been used formore than 1,000 clinical cases, became the mainstream of probes foramyloid imaging (refer to Non-Patent Document 2).

Many researchers assumed that amyloid imaging in the diagnosis ofAlzheimer's disease would be a so-called versatile diagnostic methodthat enables the diagnosis of the disease with high sensitivity andspecificity, as well as early diagnosis, differential diagnosis,diagnosis of severity (or progress), and preclinical diagnosis(so-called detection of presymptomatic high-risk individuals).

However, as clinical research progresses, issues were appearinggradually in the amyloid imaging, which was considered as versatilediagnostic method. These issues are explained by taking [¹¹C] PIB as anexample, as follows:

First, diagnosis of severity (or progress) is impossible. In otherwords, 2 years after a patient was diagnosed as Alzheimer's disease by[¹¹C] PIB, there was no change in the accumulation of the proberegardless of the progress of the clinical symptoms (refer to Non-PatentDocument 3). The reason is considered that the accumulation of Aβ towhich [¹¹C] PIB binds reaches a plateau far before MCI is seen prior todevelopment of Alzheimer's disease. Therefore, the severity or progressof Alzheimer's disease cannot be diagnosed with [¹¹C] PIB.

Second, there is a problem that considerable false positives are seen.Surprisingly, the ADNI (Alzheimer's Disease Imaging Initiative) heldahead of the International Conference on Alzheimer's Disease in Chicagoin July 2008 reported that 53% of healthy elderly were [¹¹C] PIBpositive (refer to Non-Patent Document 4). Although the incidence rateof Alzheimer's disease is considered to be 4 to 6% of the population of65 or more years old, the ADNI reported that 53% of the elderly exceptfor Alzheimer's disease patients were [¹¹C] PIB positive. Although thepresent inventors think the figure of 53% is an overestimate, thedevelopers of [¹¹C] PIB themselves recognize the possibility ofconsiderable false positives (refer to Non-Patent Document 5).

The reason for these many false positives is believed to bethat there isa considerable dispersion in the accumulation of Aβ in all of normalhealthy subjects, MCI, and Alzheimer's disease.

Furthermore, in June to July 2008, it was successively reported that theeffects of therapeutic drug (vaccines and secretase inhibitors) groups,which were expected to provide basic remedies based on the Alzheimer'sdisease/amyloid (or Aβ) hypothesis, were far below expectation. The mostshocking was the report by Holmes et al. in Lancet that Aβ vaccinescannot stop the progress of the clinical symptoms at all although Aβ wasremoved from the brain of Alzheimer's disease patients (refer toNon-Patent Document 6).

However, another important information was provided in the report inLancet; all accumulation of tau in the patients in Lancent progressed tothe final stage. FIG. 3 illustrates the Braak stage of accumulation ofAβ and tau in Alzheimer's disease. For the Braak stage of post-mortemCase 7 and 8 of the report in Lancet, Aβ was considered not to beaccumulated (or stage A), while the degree of tau accumulation was stageVI. This implies that in both cases the accumulation of Aβ was mild orless, while the accumulation of tau was the highest level of stage VI.

There were several reports that the histopathology correlated with theclinical symptoms of Alzheimer's disease was tau rather than Aβ in theearly 1990s (Non-Patent Document 7). This was unexpectedly reaffirmed bythe report by Holmes et al.

These findings strongly suggest that Aβ vaccines were less effective astherapeutic drugs after development of Alzheimer's disease and that thedegree of Aβ accumulation does not always reflect the severity ofAlzheimer's disease, as well as that it is more reasonable to trace taurather than Aβ to diagnose the severity of Alzheimer's disease.

The present inventors think that by considering the clinical outcomes ofvaccines, other therapeutic drugs, and probes for amyloid imaging, therelationship between amyloid (or Aβ) and tau in Alzheimer's diseaseshould be revised to FIG. 4. As illustrated in FIG. 4, when there is lowacccumulation of amyloid, MCI and Alzheimer's disease develop when thetau accumulation reaches the threshold, and when the accumulation ofamyloid is very high, MCI and Alzheimer's disease do not develop whenthe tau accumulation does not reach the threshold. That is to say, theamount of amyloid accumulation is not related to development of MCI andAlzheimer's disease, while tau accumulation defines this development. Itis proposed to say “amyloid (or Aβ) has no threshold, but tau has one”.

As described above, tau imaging is probably superior to amyloid imaging,in order to diagnose the severity (or progress) of Alzheimer's disease,or to detect presymptomatic high-risk individuals for Alzheimer'sdisease correctly.

The present inventors think that it is probable that “tau imaging willplay the leading role in diagnosis of Alzheimer's disease, supplementedby amyloid imaging in the future”.

The document in the relevant technical field includes, for example, (i)Okamura et al., J. Neurosci., 25 (4&), 10857-10862 (2005), (ii) EP1574500 A1, (iii) Siemens US 2010/0239496 A1, and (iv) Korea KR2010-0112423 A.

PRIOR ART DOCUMENT Patent Document

-   Non-Patent Document 1: Shoghi-Jadid K, Small G W, Agdeppa E D, Kepe    V, Ercoli L M, Siddarth P, Read S, Satyamurthy N, Petric A, Huang S    C, Barrio J R: Localization of neurofibrillary tangles and    beta-amyloid plaques in the brains of living patients with Alzheimer    disease. Am. J. Geriatr. Psychiatry 10, 24-35. 2002.-   Non-Patent Document 2: Klunk W E, Engler H, Nordberg A, Wang Y,    Blomqvist G, Holt D P, Bergstrom M, Savitcheva I, Huang G F, Estrada    S, Ausen B, Debnath M L, Barletta J, Price J C, Sandell J, Lopresti    B J, Wall A, Koivisto P, Antoni G, Mathis C A and Langstrom B.:    Imaging brain amyloid in Alzheimer's disease with Pittsburgh    Compound-B. Ann. Neurol. 55. 306-319 (2004).-   Non-Patent Document 3: Engler H, Forsberg A, Almkvist O, Blomquist    G, Larsson E, Savitcheva I, Wall A, Ringheim A, Långström B,    Nordberg A: Two-year follow-up of amyloid deposition in patients    with Alzheimer's disease. Brain. 129. 2856-2866. 2006.-   Non-Patent Document 4: Weiner: International Conference on    Alzheimer's Disease (ICAD) meeting, Chicago, 2008. July 19.-   Non-Patent Document 5: Aizenstein H J, Aizenstein H J, Nebes R D,    Saxton J A, Price J C, Mathis C A, Tsopelas N D, Ziolko S K, James J    A, Snitz B E, Houck P R, Bi W, Cohen A D, Lopresti B J, DeKosky S T,    Halligan E M, Klunk W E.: Frequent amyloid deposition without    significant cognitive impairment among the elderly. Arch Neurol. 65.    1509-1517. 2008.-   Non-Patent Document 6: Holmes C, Boche D, Wilkinson D, Yadegarfar G,    Hopkins V, Bayer A, Jones R W, Bullock R, Love S, Neal J W, Zotova    E, Nicoll J A: Long-term effects of Abeta42 immunisation in    Alzheimer's disease: follow-up of a randomised, placebo-controlled    phase I trial. Lancet. 372. 2132-2142. 2008.-   Non-Patent Document 7: Arriagada P V, Growdon J H, Hedley-Whyte E T,    Hyman B T: Neurofibrillary tangles but not senile plaques parallel    duration and severity of Alzheimer's disease. Neurology. 42.631-639.    1992.

DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention

An object of the present invention is to provide a compound which ishighly specific to tau and can image tau with satisfactory sensitivity,and also has high brain transition, low or non-recognized bone-seekingproperties and low or undetected toxicity.

Means for Solving the Problems

In light of the above problems, the present inventors have intensivelystudied and found that the compound of a formula (I), a salt thereof ora solvate thereof is a compound which is highly specific to tau and canimage tau with satisfactory sensitivity, and also has high braintransition, low or non-recognized bone-seeking properties and low ornon-recognized toxicity. They have also found that the compound of aformula (I′) can be used as a precursor of the compound of a formula(I), a salt thereof or a solvate thereof. Thus, the present inventorshave completed the present invention.

That is, the present invention provides the followings.

(1) A compound of the formula (I):

wherein

A is

R¹ is halogen, a —C(═O)-lower alkyl group (the alkyl group eachindependently may be substituted with one or more substituents selectedfrom NR^(a)R^(b), halogen and a hydroxy group), a lower alkyl group (thealkyl group each independently may be substituted with one or moresubstituents selected from halogen and a hydroxy group), a —O-loweralkyl group (the alkyl group each independently may be substituted withone or more substituents selected from halogen and a hydroxy group), or

in whichR⁴ and R⁵ each independently represents hydrogen, a lower alkyl group ora cycloalkyl group, or R⁴, R⁵ and the nitrogen atom to which they areattached are taken together to form a 3- to 8-memberednitrogen-containing aliphatic ring (one or more carbon atomsconstituting the nitrogen-containing aliphatic ring may be substitutedwith a nitrogen atom, a sulfur atom or an oxygen atom, and when a carbonatom is substituted with a nitrogen atom, the nitrogen atom may besubstituted with a lower alkyl group),or R⁴ and the nitrogen atom to which it is attached are taken togetherwith ring A to form a 8- to 16-membered nitrogen-containing fusedbicyclic ring (one or more carbon atoms constituting thenitrogen-containing fused bicyclic ring may be substituted with anitrogen atom, a sulfur atom or an oxygen atom, and when a carbon atomis substituted with a nitrogen atom, the nitrogen atom may besubstituted with a lower alkyl group) and R⁵ represents hydrogen, alower alkyl group or a cycloalkyl group,in which the line, that the dotted line intersects, means a bond of theabove general formula to the other structural moiety,R² or R³ each independently represents halogen, OH, COOH, SO₃H, NO₂, SH,NR^(a)R^(b), a lower alkyl group (the alkyl group each independently maybe substituted with one or more substituents selected from halogen and ahydroxy group) or a —O-lower alkyl group (the alkyl group eachindependently may be substituted with one or more substituents selectedfrom halogen, a hydroxy group and a —O-lower alkyl-O-lower alkyl group(the alkyl group each independently may be substituted with halogen)),ring A is unsubstituted, or substituted with R⁶ (in which R⁶ is one ormore substituents selected independently from halogen, OH, COOH, SO₃H,NO₂, SH, NR^(a)R^(b), a lower alkyl group (the alkyl group eachindependently may be substituted with one or more substituents selectedfrom halogen and a hydroxy group) and a —O-lower alkyl group (the alkylgroup each independently may be substituted with one or moresubstituents selected from halogen and a hydroxy group),R^(a) and R^(b) each independently represents hydrogen or a lower alkylgroup (the alkyl group each independently may be substituted with one ormore substituents selected from halogen and a hydroxy group),m is an integer of 0 to 4, andn is an integer of 0 to 4, or a pharmaceutically acceptable salt orsolvate thereof.(2) The compound according to (1), wherein R⁴ is halogen, a —C(═O)-loweralkyl group (the alkyl group each independently may be substituted withNH₂), a lower alkyl group (the alkyl group each independently may besubstituted with a hydroxy group), —O-lower alkyl group, or

in whichR⁴ and R⁵ each independently represents hydrogen or a lower alkyl group,ora pharmaceutically acceptable salt or solvate thereof.(3) The compound according to (1) or (2), wherein at least one of R², R³and R⁶ is a —O-lower alkyl group substituted with one hydroxy group andone halogen, or a pharmaceutically acceptable salt or solvate thereof.(4) The compound according to (3), wherein at least one of R², R³ and R⁶is represented by:

or a pharmaceutically acceptable salt or solvate thereof.(5) The compound according to (1) or (2), wherein at least one of R², R³and R⁶ is NR^(a)R^(b), and R^(a) and R^(b) each independently representshydrogen or an unsubstituted lower alkyl group, or a pharmaceuticallyacceptable salt or solvate thereof.(6) The compound according to (1), wherein the compound of the formula(I) is a compound selected from the group consisting of:

-   2-(4-aminophenyl)-8-(1-fluoromethyl-2-hydroxyethoxy)quinoline,-   2-(4-diethylaminophenyl)-6-(1-fluoromethyl-2-hydroxy)quinoline,-   2-(4-diethylaminophenyl)-7-(2-fluoromethyl-2-hydroxyethoxy)quinoline,-   2-(4-diethylaminophenyl)-8-(1-fluoromethyl-2-hydroxyethoxy)quinoline,-   2-(4-diethylaminophenyl)-7-(1-fluoromethyl-2-hydroxyethoxy)quinoline,-   2-(4-diethylaminophenyl)-4-(3-fluoro-2-hydroxypropoxy)quinoline,-   2-(4-diethylaminophenyl)-5-(1-fluoromethyl-2-hydroxyethoxy)quinoline,-   2-(4-diethylaminophenyl)-3-(1-fluoromethyl-2-hydroxyethoxy)quinoline,-   2-(4-diethylaminophenyl)-8-[(3-fluoro-2-hydroxy)propoxy]quinoline,-   2-(4-fluoromethyl-2-hydroxyethoxy)-2-(4-dimethylaminophenyl)quinoline,-   7-(1-fluoromethyl-2-hydroxyethoxy)-2-(4-methylaminophenyl)quinoline,-   2-(4-ethylmethylaminophenyl)-7-(1-fluoromethyl-2-hydroxyethoxy)-quinoline-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-dimethylaminophenyl)quinoline,-   7-[(3-fluoro-2-hydroxy)propoxy]-2-(4-methylaminophenyl)quinoline,-   7-[(3-fluoro-2-hydroxy)propoxy]-2-(4-dimethylaminophenyl)quinoline,-   2-(4-ethylmethylaminophenyl)-7-[(3-fluoro-2-hydroxy)propoxy]quinoline,-   2-(4-aminophenyl)-6-[(3-fluoro-2-hydroxy)propoxy]quinoline,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-methylaminophenyl)quinoline,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-diethylaminophenyl)quinoline,-   7-amino-2-(4-fluorophenyl)quinoline,-   2-(4-fluorophenyl)-7-dimethylaminoquinoline,-   5-amino-2-(4-fluorophenyl)quinoline,-   2-(4-fluorophenyl)-5-dimethylaminoquinoline oxalate,-   8-amino-2-(4-fluorophenyl)quinoline,-   2-(4-fluorophenyl)-8-dimethylaminoquinoline,-   6-amino-2-(4-fluorophenyl)quinoline,-   2-(4-fluorophenyl)-6-dimethylaminoquinoline,-   2-(2-aminopyrid-5-yl)-7-(1-fluoromethyl-2-hydroxyethoxy)quinoline,-   6-ethylmethylamino-2-(4-fluorophenyl)quinoline,-   6-diethylamino-2-(2-fluoropyrid-5-yl)quinoline,-   8-ethylmethylamino-2-(2-fluoropyrid-5-yl)quinoline,-   5-ethylamino-2-(2-fluoropyrid-5-yl)quinoline,-   5-diethylamino-2-(2-fluoropyrid-5-yl)quinoline,-   7-diethylamino-2-(2-fluoropyrid-5-yl)quinoline,-   7-ethylmethylamino-2-(2-fluoropyrid-5-yl)quinoline,-   2-(4-ethylaminophenyl)-6-[(3-fluoro-2-hydroxy)propoxy]quinoline,-   2-(2-aminopyrid-5-yl)-6-[(3-fluoro-2-hydroxy)propoxy]quinoline,-   2-(2-methylaminopyrid-5-yl)-6-[(3-fluoro-2-hydroxy)propoxy]quinoline,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(2-dimethylaminopyrid-5-yl)quinoline,-   2-(2-diethylaminopyrid-5-yl)-6-[(3-fluoro-2-hydroxy)propoxy]quinoline,-   2-(2-ethylaminopyrid-5-yl)-6-[(3-fluoro-2-hydroxy)propoxy]quinoline,-   1-fluoro-3-{2-[4-(4-methylpiperazin-1-yl)phenyl]quinolin-6-yloxy)propan-2-ol,-   1-fluoro-3-{2-[6-(piperazin-1-yl)pyridin-3-yl]quinolin-6-yloxy}propan-2-ol,-   1-fluoro-3-{2-[6-(4-methylpiperazin-1-yl)pyridin-3-yl]quinolin-6-yloxy}propan-2-ol,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-methyl-3,4-dihydro-2H-pyrido[3,2-b][1,4]oxazin-7-yl)quinoline,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(1-methyl-1,2,3,4-tetrahydroquinolin-6-yl)quinoline,-   6-[(3-fluoro-2-hydroxy-1,1-dimethyl)propoxy]-2-(1-methyl-1,2,3,4-tetrahydroquinolin-6-yl)quinoline,-   2-(4-amino-3-fluorophenyl)-6-dimethylaminoquinoline,-   2-[4-(amino)-3-[(3-fluoro-2-hydroxy)propoxy]phenyl]-6-methylaminoquinoline,-   2-[3-(3-fluoro-2-hydroxy-1,1-dimethyl)propoxy]-4-(dimethylamino)-phenyl]-6-dimethylaminoquinoline,-   6-amino-2-[4-(amino)-3-[(3-fluoro-2-hydroxy)propoxy]phenyl]quinoline,-   2-[3-[(3-fluoro-2-hydroxy)propoxy]-4-(dimethylamino)phenyl]-6-dimethylaminoquinoline,-   2-[3-[(3-fluoro-2-hydroxy)propoxy]-4-(methylamino)phenyl]-6-methylaminoquinoline,-   2-[4-(amino)-3-[(3-fluoro-2-hydroxy)propoxy]phenyl]-6-dimethylaminoquinoline,-   6-amino-2-[3-[(3-fluoro-2-hydroxy)propoxy]-4-(dimethylamino)phenyl]-quino    line,-   2-[3-[(3-fluoro-2-hydroxy)propoxy]-4-(dimethylamino)phenyl]-6-methylamino    quinoline,-   2-[3-[2-[2-(2-fluoroethoxy)ethoxy]ethoxy-4-(methylamino)phenyl]-6-dimethylaminoquinoline,-   2-[3-[(3-fluoro-2-hydroxy)propoxy]-4-(methylamino)phenyl]-6-dimethylamino    quinoline,-   6-amino-2-[3-[(3-fluoro-2-hydroxy)propoxy]-4-(methylamino)phenyl]-quinoline,-   2-[3-[(3-fluoro-2-hydroxy)propoxy]-2-(dimethylamino)pyrid-5-yl]-6-dimethylaminoquinoline,-   2-[3-[(3-fluoro-2-hydroxy)propoxy]-2-(dimethylamino)pyrid-5-yl]quinoline,-   6-[[2-(tetrahydro-2H-pyran-2-yloxy)-tosyloxy]propoxy]-2-(4-methyl-3,4-dihydro-2H-pyrido[3,2-b][1,4]oxazin-7-yl)quinoline,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(6-fluoropyridin-3-yl)quinoline,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-methoxyphenyl)quinoline,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-[4-(hydroxymethyl)phenyl]quinoline,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-ethanonephenyl)quinoline,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(6-methoxypyridin-3-yl)quinoline,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-ethoxyphenyl)quinoline,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-amino-3-methoxyphenyl)quinoline,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(benzamido-4-yl)quinoline,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(3-aminophenyl)quinoline, and-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(1-methyl-pyrazol-4-yl)quinoline,    or a pharmaceutically acceptable salt or solvate thereof.    (7) The compound according to any one of (1) to (6), wherein the    compound is labeled, or a pharmaceutically acceptable salt or    solvate thereof.    (8) The compound according to (7), wherein the label is a    radioactive nuclide, or a pharmaceutically acceptable salt or    solvate thereof.    (9) The compound according to (8), wherein the radioactive nuclide    is a γ-ray emitting nuclide, or a pharmaceutically acceptable salt    or solvate thereof.    (10) The compound according to (7), wherein the label is a positron    emitting nuclide, or a pharmaceutically acceptable salt or solvate    thereof.    (11) The compound according to (10), wherein the positron emitting    nuclide is selected from the group consisting of ¹¹C, ¹³N, ¹⁵O, ¹⁸F,    ³⁵mCl, ⁷⁶Br, ⁴⁵Ti, ⁴⁸V, ⁶⁰Cu, ⁶¹Cu, ⁶²Cu, ⁶⁴Cu, ⁶⁶Ga, ⁸⁹Zr, ^(94m)Tc    and ¹²⁴I, or a pharmaceutically acceptable salt or solvate thereof.    (12) The compound according to (11), wherein the positron emitting    nuclide is ¹¹C or ¹⁸F, or a pharmaceutically acceptable salt or    solvate thereof.    (13) A pharmaceutical composition comprising the compound according    to any one of (1) to (12), or a pharmaceutically acceptable salt or    solvate thereof.    (14) A pharmaceutical composition comprising the compound according    to any one of (1) to (12), or a pharmaceutically acceptable salt or    solvate thereof, and a solubilizing agent.    (15) The pharmaceutical composition according to (14), wherein the    solubilizing agent is selected from the group consisting of    Polysorbate 80, polyethylene glycol, ethanol, propylene glycol and a    combination of two or more kinds thereof.    (16) The pharmaceutical composition according to any one of (13) to    (15), which is an injection.    (17) A composition for the diagnosis of conformational disease,    comprising the compound according to any one of (1) to (12) or a    pharmaceutically acceptable salt or solvate thereof.    (18) A pharmaceutical composition for the treatment and/or    prevention of conformational disease, comprising the compound    according to any one of (1) to (12) or a pharmaceutically acceptable    salt or solvate thereof.    (19) A kit for the diagnosis of conformational disease, comprising    the compound according to any one of (1) to (12) or a    pharmaceutically acceptable salt or solvate thereof as an essential    ingredient.    (20) A composition or kit for the detection or staining of a β-sheet    structure protein, comprising the compound according to any one    of (1) to (12) or a pharmaceutically acceptable salt or solvate    thereof as an essential ingredient.    (21) The kit according to (19) or (20) for imaging diagnosis.    (22) A method of treating and/or preventing conformational disease    in a subject, which comprises administering the compound according    to any one of (1) to (12) or a pharmaceutically acceptable salt or    solvate thereof to the subject.    (23) A method of diagnosing conformational disease in a subject,    which comprises administering the compound according to any one    of (1) to (12) or a pharmaceutically acceptable salt or solvate    thereof to the subject.    (24) Use of the compound according to any one of (1) to (12) or a    pharmaceutically acceptable salt or solvate thereof for the    production of a composition or kit for the diagnosis of    conformational disease in a subject.    (25) Use of the compound according to any one of (1) to (12) or a    pharmaceutically acceptable salt or solvate thereof for the    production of a pharmaceutical composition for the treatment and/or    prevention of conformational disease in a subject.    (26) A method of detecting or staining a β-sheet structure protein    in a sample, which comprises staining the sample using the compound    according to any one of (1) to (12) or a pharmaceutically acceptable    salt or solvate thereof.    (27) Use of the compound according to any one of (1) to (12) or a    pharmaceutically acceptable salt or solvate thereof for the    production of a composition or kit for the detection or staining of    a 8-sheet structure protein.    (28) The composition, kit, method or use according to any one    of (17) to (27), wherein the conformational disease is tauopathy,    particularly Alzheimer's disease, and the 8-sheet structure protein    is tau protein.    (29) A method of producing the compound of the formula (I), which    comprises the following steps of:    (i) reacting a compound of the formula (II):

in which, R² and m are as defined in the formula (I), and R⁷ representsNH₂ or NO₂, with a compound of the formula (III):

in which A and R¹ are as defined in the formula (I), to obtain acompound of the formula (IV):

and isolating this compound as the compound of the formula (I), or (ii)optionally converting the compound of the formula (IV) into anothercompound of the formula (I) and isolating the compound.(30) A method of producing the compound of the formula (I), whichcomprises the following steps of:(i) reacting a compound of the formula (V):

in which R², R³, m and n are as defined in the formula (I), and R⁸ is ahydroxyl group or halogen, provided that at least one of R² or R³ is ahydroxy group, with a compound of the formula: OH-Ark (Ark eachindependently represents a lower alkyl group which may be substitutedwith one or more substituents selected from the group consisting ofhalogen and a hydroxy group, to obtain a compound of the formula (V′):

in which R², R³ m and n are as defined in the formula (I), and R⁸ is ahydroxyl group or halogen, provided that at least one of R² or R³ is —O—Ark (Ark is as defined above), and(ii) reacting the compound of the formula (V′) with a compound of theformula (VI) or (VII):

in which A and R¹ are as defined in the formula (I), to obtain thecompound of the formula (I) in which at least one of R² and R³ is a—O-lower alkyl group (the alkyl group each independently may besubstituted with one or more substituents selected from the groupconsisting of halogen and a hydroxy group), or isolating the compound,or(iii) optionally converting the obtained compound of the formula (I)into another compound of the formula (I), and isolating the compound.(31) A method of producing the compound of the formula (I) in which atleast one of R² and R³ is a —O-lower alkyl group (the alkyl group eachindependently may be substituted with one or more substituents selectedfrom the group consisting of halogen and a hydroxy group), whichcomprises the following steps of:(i) reacting a compound of the formula (V):

in which R², R³ m and n are as defined in the formula (I), and R⁸ is ahydroxyl group or halogen, provided that at least one of R² or R³ is ahydroxy group, with a compound of the formula (VI) or (VII):

in which A and R¹ are as defined in the formula (I), to obtain acompound of the formula (V″):

in which R¹, R², R³, A, m and n are as defined in the formula (I),provided that at least one of R² or R³ is a hydroxy group, and(ii) reacting the compound of the formula (V″) with a compound of theformula: OH-Ark (Ark each independently represents a lower alkyl groupwhich may be substituted with one or more substituents selected from thegroup consisting of halogen and a hydroxy group) to obtain compound ofthe formula (I) in which at least one of R² and R³ is a —O-lower alkylgroup (the alkyl group each independently may be substituted with one ormore substituents selected from the group consisting of halogen and ahydroxy group), and isolating the compound, or(iii) optionally converting the obtained compound of the formula (I)into another compound of the formula (I), and isolating the compound.(32) The method according to (31), wherein the compound of the formula(I) is selected from:

-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-dimethylaminophenyl)quinoline,-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-methylaminophenyl)quinoline,    and-   2-(4-ethylaminophenyl)-6-[(3-fluoro-2-hydroxy)propoxy]quinoline.    (33) A compound of the formula (I′):

in which

A is

R¹ is halogen, a —C(═O)-lower alkyl group (the alkyl group eachindependently may be substituted with one or more substituents selectedfrom NR^(a)R^(b), halogen and a hydroxy group), a lower alkyl group (thealkyl group each independently may be substituted with one or moresubstituents selected from halogen and a hydroxy group), a —O-loweralkyl group (the alkyl group each independently may be substituted withone or more substituents selected from halogen and a hydroxy group), or

in whichR⁴ and R⁵ each independently represents hydrogen, a lower alkyl group ora cycloalkyl group, or R⁴, R⁵ and the nitrogen atom to which they areattached are taken together to form a 3- to 8-memberednitrogen-containing aliphatic ring (one or more carbon atomsconstituting the nitrogen-containing aliphatic ring may be substitutedwith a nitrogen atom, a sulfur atom or an oxygen atom, and when a carbonatom is substituted with a nitrogen atom, the nitrogen atom may besubstituted with a lower alkyl group), or R⁴ and the nitrogen atom towhich it is attached are taken together with ring A to form a 8- to16-membered nitrogen-containing fused bicyclic ring (one or more carbonatoms constituting the nitrogen-containing fused bicyclic ring may besubstituted with a nitrogen atom, a sulfur atom or an oxygen atom, andwhen a carbon atom is substituted with a nitrogen atom, the nitrogenatom may be substituted with a lower alkyl group), R⁵ is hydrogen, alower alkyl group or a cycloalkyl group, in which the line, that thedotted line intersects, means a bond of the above general formula to theother structural moiety, R² or R³ each independently may be substitutedwith halogen, OH, COOH, SO₃H, NO₂, SH, NR^(a)R^(b), a lower alkyl group(the lower alkyl group each independently may be substituted with one ormore substituents selected from a p-toluenesulfonyloxy group (tosyloxygroup, TsO), a methanesulfonyloxy group, a trifluoromethanesulfonyloxygroup or a 2-tetrahydropyranyloxy (OTHP), halogen and a hydroxy group)or a —O-lower alkyl group (the lower alkyl group is substituted with ap-toluenesulfonyloxy group (tosyloxy group, TsO), a methanesulfonyloxygroup, trifluoromethanesulfonyloxy group or a 2-tetrahydropyranyloxy(OTHP), and also may be substituted with a hydroxy group),ring A is unsubstituted or substituted with R⁶ (in which R⁶ is one ormore substituents selected independently from halogen, OH, COOH, SO₃H,NO₂, SH, NR^(a)R^(b), a lower alkyl group (the lower alkyl group eachindependently may be substituted with one or more substituents selectedfrom the group consisting of a p-toluenesulfonyloxy group (tosyloxygroup, TsO), a methanesulfonyloxy group, a trifluoromethanesulfonyloxygroup or a 2-tetrahydropyranyloxy (OTHP), halogen and a hydroxy group)and an —O-lower alkyl group (the alkyl group each independently may besubstituted with one or more substituents selected from ap-toluenesulfonyloxy group (tosyloxy group, TsO), a methanesulfonyloxygroup, a trifluoromethanesulfonyloxy group or a 2-tetrahydropyranyloxy(OTHP), halogen, a hydroxy group and an —O-lower alkyl group-O-loweralkyl group (the alkyl group each independently may be substituted withhalogen))), R^(a) and R^(b) independently represents hydrogen or a loweralkyl group (the alkyl group and the lower alkyl group eachindependently may be substituted with one or more substituents selectedfrom a p-toluenesulfonyloxy group (tosyloxy group, TsO), amethanesulfonyloxy group, a trifluoromethanesulfonyloxy group or a2-tetrahydropyranyloxy (OTHP), halogen and a hydroxy group),m is an integer of 0 to 4, andn is an integer of 0 to 4,provided that at least one of R², R³ and R⁶ represents an —O-lower alkylgroup (the lower alkyl group is substituted with a p-toluenesulfonyloxygroup (tosyloxy group, TsO), a methanesulfonyloxy group, atrifluoromethanesulfonyloxy group or a 2-tetrahydropyranyloxy (OTHP),and may also be substituted with one or more substituents selected fromhalogen and a hydroxy group), or a pharmaceutically acceptable salt orsolvate thereof.(34) The compound according to (33), wherein at least one of R², R³ andR⁶ is a group of the formula:

(35) The compound according to (33), wherein the compound of the formula(I′) is a compound selected from the group consisting of:

-   2-(4-diethylaminophenyl)-6-[(2-hydroxy-1-tosyloxymethyl)ethoxy]quinoline,-   2-(4-aminophenyl)-8-(2-hydroxy-1-tosyloxymethylethoxy)quinoline,-   2-(4-diethylaminophenyl)-8-(2-hydroxy-1-tosyloxymethylethoxy)quinoline,-   2-(4-diethylaminophenyl)-8-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   2-(4-diethylaminophenyl)-7-(2-hydroxy-1-tosyloxymethylethoxy)quinoline,-   2-(4-diethylaminophenyl)-7-[[(2-tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   7-(2-hydroxy-1-tosyloxymethylethoxy)-2-(4-dimethylaminophenyl)quinoline,-   7-(2-hydroxy-1-tosyloxymethylethoxy)-2-(4-methylaminophenyl)quinoline,-   2-(4-ethylmethylaminophenyl)-7-(2-hydroxy-1-tosyloxymethylethoxy)quinoline,-   2-(4-diethylaminophenyl)-5-(2-hydroxy-1-tosyloxymethylethoxy)quinoline,-   2-(4-ethylmethylaminophenyl)-7-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   2-(4-methylaminophenyl)-7-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   2-(4-dimethylaminophenyl)-7-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   2-(4-methylaminophenyl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   2-(4-diethylaminophenyl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   2-(4-dimethylaminophenyl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   2-(4-aminophenyl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   2-(4-ethylaminophenyl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   2-(2-aminopyrid-5-yl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   2-(2-methylaminopyrid-5-yl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   2-(2-dimethylaminopyrid-5-yl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   2-(2-diethylaminopyrid-5-yl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   2-(2-ethylaminopyrid-5-yl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline,-   2-[4-(methylamino)-3-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]phenyl]-6-dimethylaminoquinoline,    and-   2-[4-(dimethylamino)-3-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]phenyl]-6-dimethylaminoquinoline.    (36) A kit for producing the labeled compound according to any one    of (33) to (35), or a pharmaceutically acceptable salt or solvate    thereof, comprising: the compound according to any one of (33) to    (35), or a pharmaceutically acceptable salt or solvate thereof,    a labeling agent, and    optionally, instructions for carrying out labeling.    (37) The kit according to (36), wherein the labeling agent is a    radioactive nuclide.    (38) The kit according to (37), wherein the radioactive nuclide is a    γ-ray emitting nuclide.    (39) The kit according to (36), wherein the labeling agent is a    positron emitting nuclide.    (40) The kit according to (39), wherein the positron emitting    nuclide is selected from the group consisting of ¹¹C, ¹³N, ¹⁵O, ¹⁸F,    ³⁵mCl, ⁷⁶Br, ⁴⁵Ti, ⁴⁸V, ⁶⁰Cu, ⁶¹Cu, ⁶²Cu, ⁶⁴Cu, ⁶⁶Ga, ⁸⁹Zr, ^(94m)Tc    and ¹²⁴I.    (41) The kit according to (40), wherein the positron emitting    nuclide is ¹¹C or ¹⁸F.    (42) A method for producing the compound according to (7), which    comprises the step of reacting the compound according to (34) with a    labeling agent.    (43) The method according to (42), wherein the labeling agent is a    radioactive nuclide.

Effects of the Invention

According to the present invention, there is provided a compound havingvery high safety, which is highly specific to tau and can image tau withsatisfactory sensitivity, and also has high brain transition, low orundetected bone-seeking properties and low or undetected toxicity, and aprecursor thereof. Accordingly, the diagnosis, the treatment and/orprevention of tauopathy can be carried out using the compound of thepresent invention. Also, according to the present invention, it becomespossible to carry out imaging diagnosis of tauopathy, particularlyimaging diagnosis using PET. Accordingly, according to the presentinvention, it becomes possible to carry out accurate diagnosis,effective treatment and prevention in the early stages of tauopathy,particularly Alzheimer's disease.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a diagram showing deviation between a clinical picture and ahistological picture in Alzheimer's disease. Cited from Alzheimer'sDisease (written by Yasuo IHARA, Hiroyuki ARAKI), Asahi Shimbun Company,2007, Tokyo, partly revised. In the onset of Alzheimer's disease ofpatients aged 80 years, accumulation of Aβ starts at the age of 50 yearsand has already reach a plateau at the age of 60 years. On the otherhand, tau accumulation proceeds age-dependently at the age of 70 years.

FIG. 2 illustrates PET probes for amyloid imaging, which have beendeveloped so far.

FIG. 3 is a diagram showing stages of Aβ accumulation and tauaccumulation in Alzheimer's disease. Cited from Braak & Braak: Neurobiolaging. 18.351-357. 1997, partly revised. Referring to stages of Braakafter death of cases 7 and 8 in Non-Patent Document 6, Aβ accumulationwas considered as Cases devoid of amyloid (or stage A), while tauaccumulation was in stage VI. That is, this means that although Aβaccumulation is mild or globally mild in both cases, tau accumulationwas in stage VI in which an accumulation level is the highest.

FIG. 4 is a graph showing a relation between amyloid (or Aβ) and tau inAlzheimer's disease (proposed by the present inventors). As shown inupper, middle and lower columns, when amyloid is not much accumulated,MCI and Alzheimer's disease develop when the tau accumulation reachesthe threshold, and when amyloid is strongly accumulated, MCI andAlzheimer's disease do not develop when the tau accumulation does notreach the threshold. That is to say, the amount of amyloid accumulationis not related to development of MCI and Alzheimer's disease, while tauaccumulation defines this development. In other words, “amyloid (or Aβ)has no threshold, but tau has one.

The upper panel of FIG. 5 is a THK-5035 stained image in a brain sectionof Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 5 is a THK-5038 stainedimage in a brain section of Alzheimer's disease patients. The outlinearrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 6 is a THK-5058 stained image in a brain sectionof Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 6 is a THK-5064 stainedimage in a brain section of Alzheimer's disease patients. The outlinearrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 7 is a THK-5065 stained image in a brain sectionof Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 7 is a THK-5066 stainedimage in a brain section of Alzheimer's disease patients. The outlinearrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 8 is a THK-5071 stained image in a brain sectionof Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 8 is a THK-5077 stainedimage in a brain section of Alzheimer's disease patients. The outlinearrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 9 is a THK-5078 stained image in a brain sectionof Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 9 is a THK-5079 stainedimage in a brain section of Alzheimer's disease patients. The outlinearrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 10 is a THK-5080 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 10 is lower panel is aTHK-5081 stained image in a brain section of Alzheimer's diseasepatients. The outline arrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 11 is a THK-5082 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 11 is a THK-5087stained image in a brain section of Alzheimer's disease patients. Theoutline arrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 12 is a THK-5088 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 12 is a THK-5089stained image in a brain section of Alzheimer's disease patients. Theoutline arrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 13 is a THK-5091 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 13 is a THK-5092stained image in a brain section of Alzheimer's disease patients. Theoutline arrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 14 is a THK-5097 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 10 is a THK-5098stained image in a brain section of Alzheimer's disease patients. Theoutline arrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 15 is a THK-5059 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 15 is a THK-5075stained image in a brain section of Alzheimer's disease patients. Theoutline arrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 16 is a THK-5076 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 15 is a THK-5086stained image in a brain section of Alzheimer's disease patients. Theoutline arrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 17 is a THK-5100 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 17 is a THK-5105stained image in a brain section of Alzheimer's disease patients. Theoutline arrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 18 is a THK-5106 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 18 is a THK-5107stained image in a brain section of Alzheimer's disease patients. Theoutline arrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 19 is a THK-5112 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 19 is a THK-5116stained image in a brain section of Alzheimer's disease patients. Theoutline arrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 20 is a THK-5117 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 20 is a THK-932 stainedimage in a brain section of Alzheimer's disease patients. The outlinearrowhead denotes neurofibrillary tangles.

FIG. 21 shows an autoradiography image (upper left-hand and upperright-hand, respectively) of [¹⁸F] BF-227 and [¹⁸F] THK-5035, athioflavin S (TF-S) stained image (lower left-hand) in a serial section,and an anti-Tau antibody (Tau) stained image (lower right-hand).

FIG. 22 shows an image of [¹⁸F] THK-5105 autoradiography (upper left)and an image of anti-phosphorylated tau antibody (pTau) staining, ofsections of the hippocampus from an Alzheimer's disease patient (afemale of 83 years old and with a brain weight of 900 g). The lowerpanels show, from left to right, higher magnification images of [¹⁸F]THK-5105 autoradiography, of anti-phosphorylated tau antibody (pTau)staining, and of unlabeled THK-5105 staining, with insets representing amuch higher magnification of the anti-phosphorylated tau antibody (pTau)staining and of the unlabeled THK-5105 staining.

FIG. 23 shows images of [¹⁸F] THK-5105, [¹⁸F] THK-5117, and [¹⁸F]THK-5125 autoradiography of sections of the lateral temporal cortex andof the medial temporal cortex from an Alzheimer's disease patient (afemale of 77 years old and with a brain weight of 1,100 g).

FIG. 24 shows binding affinities (Kis) of various probes to a tau. Thetau used was aggregates of mutated tau (K18-ΔK280) and the radioactiveligand used was [¹⁸F] THK-5105.

The upper panel of FIG. 25 is a THK-5136 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 25 is a THK-5153stained image in a brain section of Alzheimer's disease patients. Theoutline arrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 26 is a THK-5157 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 26 is a THK-5128stained image in a brain section of Alzheimer's disease patients. Theoutline arrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 27 is a THK-5147 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles.

The upper panel of FIG. 28 is a THK-5155 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 28 is a THK-5156stained image in a brain section of Alzheimer's disease patients. Theoutline arrowhead denotes neurofibrillary tangles.

The upper panel of FIG. 29 is a THK-5164 stained image in a brainsection of Alzheimer's disease patients. The outline arrowhead denotesneurofibrillary tangles. The lower panel of FIG. 29 is a THK-5154stained image in a brain section of Alzheimer's disease patients. Theoutline arrowhead denotes neurofibrillary tangles.

MODE FOR CARRYING OUT THE INVENTION

The compounds of the present invention are compounds of formulae (I) and(I′) described below, or salts or solvates thereof. As used herein,“compound of the present invention” and “compound according to thepresent invention” include the compounds of formulae (I) and (I′)described below, and salts and solvates thereof, unless otherwisespecified.

As used herein, “lower alkyl group” means a linear or branched alkylgroup having 1 to 6 carbon atoms, and specific examples thereof includea methyl group, an ethyl group, a propyl group, an isopropyl group, abutyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, apentyl group, an isoamyl group, a neopentyl group, an isopentyl group, a1,1-dimethylpropyl group, a 1-methylbutyl group, a 2-methylbutyl group,a 1,2-dimethylpropyl group, a hexyl group, an isohexyl group, a1-methylpentyl group, a 2-methylpentyl group, a 3-methylpentyl group, a1,1-dimethylbutyl group, a 1,2-dimethylbutyl group, a 2,2-dimethylbutylgroup, a 1,3-dimethylbutyl group, a 2,3-dimethylbutyl group, a3,3-dimethylbutyl group, a 1-ethylbutyl group, a 2-ethylbutyl group, a1,2,2-trimethylpropyl group, a 1-ethyl-2-methylpropyl group and thelike. The term “lower alkoxy” means —O-lower alkyl.

As used herein, “cycloalkyl group” means a cycloalkyl group having 3 to7 carbon atoms, and specific examples thereof include a cyclopropylgroup, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group and acycloheptyl group.

As used herein, “halogen” means fluorine, chlorine, bromine or iodine.

As used herein, “tau protein” and “tau” have the same meanings. As usedherein, “amyloidbeta protein”, “amyloid β protein”, “Aβ protein”,“amyloidbeta”, “amyloid β” and “Aβ” have the same meanings.

In case an asymmetric carbon atom exists in the compound of the presentinvention, a mixture of isomers, and individual isomer are also includedin the compound of the present invention.

For example, in case one asymmetric carbon exists in the compound of thepresent invention, each optically active compound can be separatelysynthesized, or individual optical isomer can be separated by columnchromatography. For example, in case the optical isomer is separated bycolumn chromatography, a column to be used includes, for example,CHIRALPAK AD (manufactured by DAICEL CHEMICAL INDUSTRIES, LTD.) or thelike. Also, a solvent used in column chromatography may be a solventwhich is usually used to separate the isomer. For example, chloroform,acetonitrile, ethyl acetate, methanol, ethanol, acetone, hexane, waterand the like are used alone, or two or more kinds of these solvents canalso be used in combination.

In order to disclose the compound of the formula (I):

[wherein the respective symbols are as defined above] according to thepresent invention in more specifically, various symbols used in theformula (I) will be described by way of specific examples.Ring A means

in which the line, that the dotted line intersects, means a bond of theabove general formula to the other structural moiety. That is, bondsexisting at 2- and 5-positions of pyridine ring are respectivelyattached to R¹ and quinoline ring of the general formula (I). The ring Ais unsubstituted or substituted with one to four substituents, andpreferably unsubstituted or substituted with one substituent selectedfrom fluorine, (3-fluoro-2-hydroxy)propoxy,(3-fluoro-2-hydroxy-1,1-dimethyl)propoxy,2-[2-(2-fluoroethoxy)ethoxy]ethoxy and methoxy.R¹ is halogen, a —C(═O)-lower alkyl group (the alkyl group eachindependently may be substituted with one or more substituents selectedfrom NR^(a)R^(b), halogen and a hydroxy group), a lower alkyl group (thealkyl group each independently may be substituted with one or moresubstituents selected from halogen and a hydroxy group), a —O-loweralkyl group (the alkyl group each independently may be substituted withone or more substituents selected from halogen and a hydroxy group), or

in whichR⁴ and R⁵ each independently represents hydrogen, a lower alkyl group ora cycloalkyl group, or R⁴, R⁵ and the nitrogen atom to which they areattached are taken together to form a 3- to 8-memberednitrogen-containing aliphatic ring (one or more carbon atomsconstituting the nitrogen-containing aliphatic ring may be substitutedwith a nitrogen atom, a sulfur atom or an oxygen atom, and when a carbonatom is substituted with a nitrogen atom, the nitrogen atom may besubstituted with a lower alkyl group), or R⁴ and the nitrogen atom towhich it is attached are taken together with ring A to form a 8- to16-membered nitrogen-containing fused bicyclic ring (one or more carbonatoms constituting the nitrogen-containing fused bicyclic ring may besubstituted with a nitrogen atom, a sulfur atom or an oxygen atom, andwhen a carbon atom is nitrogen atom a carbon atom is substituted with anitrogen atom, the nitrogen atom may be substituted with a lower alkylgroup), R⁵ is hydrogen, a lower alkyl group or a cycloalkyl group.

The “lower alkyl group” represented by R⁴ and R⁵ means the same groupsas those in the lower alkyl group defined above. Among these groups, amethyl group, an ethyl group and a propyl group are preferable, and amethyl group is more preferable.

The “cycloalkyl group” represented by R⁴ and R⁵ means the same groups asthose in the cycloalkyl group defined above.

Specific examples of the 3- to 8-membered nitrogen-containing aliphaticring formed by taking R⁴, R⁵ and the nitrogen atom to which they areattached together (carbon atoms constituting the nitrogen-containingaliphatic ring may be substituted with a nitrogen atom, a sulfur atom oran oxygen atom and, in case carbon atoms are substituted with a nitrogenatom, the nitrogen atom may be substituted with a lower alkyl group)include groups of formula:

wherein Z is O, S, CH₂ or NR^(e), and R^(e) represents hydrogen or aC₁₋₄ alkyl group. Among these groups, a morpholino group, a piperazinegroup and a 4-methyl-piperazine group are preferable.

Specific examples of the 8- to 16-membered nitrogen-containing fusedbicyclic ring formed by taking R⁴ and the nitrogen atom to which it isattached together with ring A (one or more carbon atoms constituting thenitrogen-containing fused bicyclic ring may be substituted with anitrogen atom, a sulfur atom or an oxygen atom and, incase carbon atomsare substituted with a nitrogen atom, the nitrogen atom may besubstituted with a lower alkyl group) include groups of formula:

wherein Z is O, S, CH₂ or NR^(e), and R^(e) represents hydrogen or aC₁₋₄ alkyl group. Among these groups,

is particularly preferable.

R², R³ and R⁶ each independently represents halogen, OH, COOH, SO₃H,NO₂, SH, NR^(a)R^(b) or a lower alkyl group (the alkyl group issubstituted with a halogen atom, and also may be substituted with ahydroxy group) or a —O-lower alkyl group (the alkyl group eachindependently may be substituted with one or more substituents selectedfrom the group consisting of halogen, a hydroxy group and an —O-loweralkyl group-O-lower alkyl group (the alkyl group each independently maybe substituted with halogen)).

At least one of R², R³ and R⁶ is preferably a —O-lower alkyl group (thealkyl group is substituted with a halogen atom, and also may besubstituted with a hydroxy group). Among these groups, a —O-lower alkylgroup substituted with a halogen atom or a —O-lower alkyl groupsubstituted with a halogen atom and a hydroxy group is preferable, and agroup of a formula:

is more preferable.

R^(a) and R^(b) independently represents hydrogen or a lower alkyl group(the alkyl group is substituted with a halogen atom, and also may besubstituted with a hydroxy group). Preferred R^(a) and R^(b) arehydrogens.

m is an integer of 0 to 4, and preferably 1.

n is an integer of 0 to 4. Preferably, all R⁴(s) are hydrogens.

Examples of preferred compounds of formula (I) include:

-   2-(4-aminophenyl)-8-(1-fluoromethyl-2-hydroxyethoxy)quinoline    (THK-5004),-   2-(4-diethylaminophenyl)-6-(1-fluoromethyl-2-hydroxy)quinoline    (THK-5035),-   2-(4-diethylaminophenyl)-7-(2-fluoromethyl-2-hydroxyethoxy)quinoline    (THK-5038),-   2-(4-diethylaminophenyl)-8-(1-fluoromethyl-2-hydroxyethoxy)quinoline    (THK-5051),-   2-(4-diethylaminophenyl)-7-(1-fluoromethyl-2-hydroxyethoxy)quinoline    (THK-5058),-   2-(4-diethylaminophenyl)-4-(3-fluoro-2-hydroxypropoxy)quinoline    (THK-5059),-   2-(4-diethylaminophenyl)-5-(1-fluoromethyl-2-hydroxyethoxy)quinoline    (THK-5064),-   2-(4-diethylaminophenyl)-3-(1-fluoromethyl-2-hydroxyethoxy)quinoline    (THK-5065),-   2-(4-diethylaminophenyl)-8-[(3-fluoro-2-hydroxy)propoxy]quinoline    (THK-5066),-   2-(4-fluoromethyl-2-hydroxyethoxy)-2-(4-dimethylaminophenyl)quinoline    (THK-5071),-   7-(1-fluoromethyl-2-hydroxyethoxy)-2-(4-methylaminophenyl)quinoline    (THK-5077),-   2-(4-ethylmethylaminophenyl)-7-(1-fluoromethyl-2-hydroxyethoxy)-quinoline    (THK-5078),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-dimethylaminophenyl)quinoline    (THK-5105),-   7-[(3-fluoro-2-hydroxy)propoxy]-2-(4-methylaminophenyl)quinoline    (THK-5106),-   7-[(3-fluoro-2-hydroxy)propoxy]-2-(4-dimethylaminophenyl)quinoline    (THK5107),-   2-(4-ethylmethylaminophenyl)-7-[(3-fluoro-2-hydroxy)propoxy]quinoline    (THK-5112),-   2-(4-aminophenyl)-6-[(3-fluoro-2-hydroxy)propoxy]quinoline    (THK-5116),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-methylaminophenyl)quinoline    (THK-5117),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-diethylaminophenyl)quinoline    (THK-5122),-   7-amino-2-(4-fluorophenyl)quinoline (THK-5075),-   2-(4-fluorophenyl)-7-dimethylaminoquinoline (THK-5076),-   5-amino-2-(4-fluorophenyl)quinoline (THK-5079),-   2-(4-fluorophenyl)-5-dimethylaminoquinoline oxalate (THK-5080),-   8-amino-2-(4-fluorophenyl)quinoline (THK-5081),-   2-(4-fluorophenyl)-8-dimethylaminoquinoline (THK-5082),-   6-amino-2-(4-fluorophenyl)quinoline (THK-5086),-   2-(4-fluorophenyl)-6-dimethylaminoquinoline (THK-5087),-   2-(2-aminopyrid-5-yl)-7-(1-fluoromethyl-2-hydroxyethoxy)quinoline    (THK-932),-   6-ethylmethylamino-2-(4-fluorophenyl)quinoline (THK-5100),-   6-diethylamino-2-(2-fluoropyrid-5-yl)quinoline (THK-5088),-   8-ethylmethylamino-2-(2-fluoropyrid-5-yl)quinoline (THK-5089),-   5-ethylamino-2-(2-fluoropyrid-5-yl)quinoline (THK-5091),-   5-diethylamino-2-(2-fluoropyrid-5-yl)quinoline (THK-5092),-   7-diethylamino-2-(2-fluoropyrid-5-yl)quinoline (THK-5097),-   7-ethylmethylamino-2-(2-fluoropyrid-5-yl)quinoline (THK-5098),-   2-(4-ethylaminophenyl)-6-[(3-fluoro-2-hydroxy)propoxy]quinoline    (THK-5125),-   2-(2-aminopyrid-5-yl)-6-[(3-fluoro-2-hydroxy)propoxy]quinoline    (THK-5127),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(2-dimethylaminopyrid-5-yl)quinoline    (THK-5129),-   2-(2-diethylaminopyrid-5-yl)-6-[(3-fluoro-2-hydroxy)propoxy]quinoline    (THK-5130),-   2-(2-ethylaminopyrid-5-yl)-6-[(3-fluoro-2-hydroxy)propoxy]quinoline    (THK-5142),-   2-(2-methylaminopyrid-5-yl)-6-[(3-fluoro-2-hydroxy)propoxy]quinoline    (THK-5151),-   1-fluoro-3-{2-[4-(4-methylpiperazin-1-yl)phenyl]quinolin-6-yloxy)propan-2-ol    (THK-5177),-   1-fluoro-3-{2-[6-(piperazin-1-yl)pyridin-3-yl]quinolin-6-yloxy}propan-2-ol    (THK-5178),-   1-fluoro-3-{2-[6-(4-methylpiperazin-1-yl)pyridin-3-yl]quinolin-6-yloxy}propan-2-ol    (THK-5180),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-methyl-3,4-dihydro-2H-pyrido[3,2-b][1,4]oxazin-7-yl)quinoline    (THK-5136),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(1-methyl-1,2,3,4-tetrahydroquinolin-6-yl)quinoline    (THK-5153),-   6-[(3-fluoro-2-hydroxy-1,1-dimethyl)propoxy]-2-(1-methyl-1,2,3,4-tetrahydroquinolin-6-yl)quinoline    (THK-5157),-   2-(4-amino-3-fluorophenyl)-6-dimethylaminoquinoline (THK-5128),-   2-[4-(amino)-3-[(3-fluoro-2-hydroxy)propoxy]phenyl]-6-methylaminoquinoline    (THK-5147),-   2-[3-(3-fluoro-2-hydroxy-1,1-dimethyl)propoxy]-4-(dimethylamino)-phenyl]-6-dimethylaminoquinoline    (THK-5148),-   6-amino-2-[4-(amino)-3-[(3-fluoro-2-hydroxy)propoxy]phenyl]quinoline    (THK-5155),-   2-[3-[(3-fluoro-2-hydroxy)propoxy]-4-(dimethylamino)phenyl]-6-dimethylaminoquinoline    (THK-5156),-   2-[3-[(3-fluoro-2-hydroxy)propoxy]-4-(methylamino)phenyl]-6-methylaminoquinoline    (THK-5158),-   2-[4-(amino)-3-[(3-fluoro-2-hydroxy)propoxy]phenyl]-6-dimethylaminoquinoline    (THK-5159),-   6-amino-2-[3-[(3-fluoro-2-hydroxy)propoxy]-4-(dimethylamino)phenyl]-quino    line (THK-5160),-   2-[3-[(3-fluoro-2-hydroxy)propoxy]-4-(dimethylamino)phenyl]-6-methylamino    quinoline (THK-5161),-   2-[3-[2-[2-(2-fluoroethoxy)ethoxy]ethoxy-4-(methylamino)phenyl]-6-dimethylaminoquinoline    (THK-5162),-   2-[3-[(3-fluoro-2-hydroxy)propoxy]-4-(methylamino)phenyl]-6-dimethylaminoquinoline    (THK-5164),-   6-amino-2-[3-[(3-fluoro-2-hydroxy)propoxy]-4-(methylamino)phenyl]-quinoline    (THK-5165),-   2-[3-[(3-fluoro-2-hydroxy)propoxy]-2-(dimethylamino)pyrid-5-yl]-6-dimethylaminoquinoline    (THK-5154),-   2-[3-[(3-fluoro-2-hydroxy)propoxy]-2-(dimethylamino)pyrid-5-yl]quinoline    (THK-5166),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(6-fluoropyridin-3-yl)quinoline    (THK-5170),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-methoxyphenyl)quinoline    (THK-5171),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-[4-(hydroxymethyl)phenyl]quinoline    (THK-5172),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-ethanonephenyl)quinoline    (THK-5173),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(6-methoxypyridin-3-yl)quinoline    (THK-5174),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-ethoxyphenyl)quinoline    (THK-5175),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-amino-3-methoxyphenyl)quinoline    (THK-5176),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(benzamido-4-yl)quinoline    (THK-5179),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(3-aminophenyl)quinoline    (THK-5181), and-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(1-methyl-pyrazol-4-yl)quinoline    (THK-5182).

Examples of more preferred compound of the formula (I) include:

-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-dimethylaminophenyl)quinoline    (THK-5105),-   6-[(3-fluoro-2-hydroxy)propoxy]-2-(4-methylaminophenyl)quinoline    (THK-5117), and-   2-(4-ethylaminophenyl)-6-[(3-fluoro-2-hydroxy)propoxy]quinoline    (THK-5125).

As shown in Examples, the compound of the formula (I) is highly specificto tau, and also has high brain uptake. Also, the compound of theformula (I) is a compound having very high safety, which has low ornon-recognized bone-seeking properties and low or non-recognizedtoxicity. Accordingly, the diagnosis of tauopathy can be carried outusing the compound of formula (I) as a probe against tau, and also thetreatment and/or prevention of tauopathy can be carried out by using thecompound of the formula (I). Particularly, the compound of formula (I)is suited for imaging diagnosis of tauopathy, particularly imagingdiagnosis using PET. Accordingly, it becomes possible to carry outaccurate diagnosis, effective treatment and prevention in the earlystages of tauopathy, particularly Alzheimer's disease, using thecompound of formula (I).

The conformational disease is the disease in which a protein having aspecific β-sheet structure accumulates, and there are various diseasescharacterized by deposition of an insoluble fibrillar protein to variousinternal organs and tissues. These diseases include Alzheimer's disease,prion disease, dementia with Lewy bodies, Parkinson's disease,Huntington's disease, spinal and bulbar atrophy,dentate-rubro-pallido-luysian atrophy, Spinocerebellar Degeneration,Machado-Joseph Disease, Amyophic Lateral Sclerosis (ALS), Down'ssyndrome, Pick's disease, FTDP-17 (Frontotemporal Dementia andParkinsonism linked to Chromosome 17), LNTD (Limbic Neurofibrillarytangles Demetia), Sudanophiloc Leukodystrophy, amyloidosis and the like.

In the present invention, the conformational disease preferably meansdisease (tauopathy) having a cardinal symptom such as intracerebralaccumulation of tau protein. Tauopathy includes Alzheimer's disease,Pick's disease, progressive supranuclear palsy (PSP) and the like.

In order to disclose the compound of a formula (I′):

[wherein the respective symbols are as defined above] which is aprecursor of the compound of the formula (I) according to the presentinvention in more specifically, various symbols used in the formula (I′)will be described by way of specific examples.

A and R¹ are the same as defined in the formula (I), as described above.R^(a) and R^(b) are also the same as defined in the formula (I), asdescribed above.

R², R³ and R⁶ each independently represents halogen, —OH, —COOH, —SO₃H,—NO₂, —SH, —NR^(a)R^(b) (R^(a) and R^(b) independently representshydrogen or a lower alkyl group (the alkyl group each independently maybe substituted with one or more substituents selected from halogen and ahydroxy group)), a lower alkyl group (the lower alkyl group eachindependently may be substituted with one or more substituents selectedfrom a p-toluenesulfonyloxy group (tosyloxy group, TsO), amethanesulfonyloxy group, a trifluoromethanesulfonyloxy group or a2-tetrahydropyranyloxy (OTHP), halogen and a hydroxy group), a —O-loweralkyl group (the lower alkyl group each independently maybe substitutedwith one or more substituents selected from a p-toluenesulfonyloxy group(tosyloxy group, TsO), a methanesulfonyloxy group, atrifluoromethanesulfonyloxy group or a 2-tetrahydropyranyloxy (OTHP),halogen, a hydroxy group and a —O-lower alkyl group-O-lower alkyl group(the alkyl group each independently may be substituted with halogen)),in which at least one of R², R³ and R⁶ is a —O-lower alkyl group (thelower alkyl group is substituted with a p-toluenesulfonyloxy group(tosyloxy group, TsO), a methanesulfonyloxy group, atrifluoromethanesulfonyloxy group or a 2-tetrahydropyranyloxy (OTHP),and also may be substituted with one or more substituents selected fromhalogen, hydroxy group and a —O-lower alkyl group-O-lower alkyl group(the alkyl group each independently may be substituted with halogen)).

At least one of R², R³ and R⁶ is preferably a group of a formula:

m is an integer of 0 to 4. Preferably, m is 0.

n is an integer of 0 to 4. Preferably, n is 0.

Examples of preferred compound of the formula (I′) include:

-   2-(4-diethylaminophenyl)-6-[(2-hydroxy-1-tosyloxymethyl)ethoxy]quinoline    (THK-5039),-   2-(4-aminophenyl)-8-(2-hydroxy-1-tosyloxymethylethoxy)quinoline    (THK-5041),-   2-(4-diethylaminophenyl)-8-(2-hydroxy-1-tosyloxymethylethoxy)    quinoline (THK-5050),-   2-(4-diethylaminophenyl)-8-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline    (THK-5070),-   2-(4-diethylaminophenyl)-7-(2-hydroxy-1-tosyloxymethylethoxy)    quinoline (THK-5072),-   2-(4-diethylaminophenyl)-7-[[(2-tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline    (THK-5073),-   7-(2-hydroxy-1-tosyloxymethylethoxy)-2-(4-dimethylaminophenyl)    quinoline (THK-5090),-   7-(2-hydroxy-1-tosyloxymethylethoxy)-2-(4-methylaminophenyl)    quinoline (THK-5095),-   2-(4-ethylmethylaminophenyl)-7-(2-hydroxy-1-tosyloxymethylethoxy)quinoline    (THK-5096),-   2-(4-diethylaminophenyl)-5-(2-hydroxy-1-tosyloxymethylethoxy)    quinoline (THK-5099),-   2-(4-ethylmethylaminophenyl)-7-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline    (THK-5111),-   2-(4-methylaminophenyl)-7-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline    (THK-5113),-   2-(4-dimethylaminophenyl)-7-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline    (THK-5115),-   2-(4-methylaminophenyl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline    (THK-5119),-   2-(4-diethylaminophenyl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline    (THK-5120),-   2-(4-dimethylaminophenyl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline    (THK-5121),-   2-(4-aminophenyl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]-propoxy]quinoline    (THK-5123),-   2-(4-ethylaminophenyl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline    (THK-5131),-   2-(2-aminopyrid-5-yl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline    (THK-5150),-   2-(2-methylaminopyrid-5-yl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline    (THK-5152),-   2-(2-dimethylaminopyrid-5-yl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline    (THK-5135),-   2-(2-diethylaminopyrid-5-yl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline    (THK-5138),-   2-(2-ethylaminopyrid-5-yl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline    (THK-5143),-   2-[4-(methylamino)-3-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]phenyl]-6-dimethylaminoquinoline    (THK-5163),-   6-[[2-(tetrahydro-2H-pyran-2-yloxy)-tosyloxy]propoxy]-2-(4-methyl-3,4-dihydro-2H-pyrido[3,2-b][1,4]oxazin-7-yl)quinoline    (THK-5167), and-   2-[4-(dimethylamino)-3-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy]phenyl]-6-dimethylaminoquinoline    (THK-5168).

The compound of the formula (I′) can be used as a synthetic precursor ofthe compound of the formula (I). The method of converting into thecompound of the formula (I) from the compound of the formula (I′) iswell known to a person with an ordinary skill in the art, and thecompound of the formula (I) can be easily obtained.

Salts of the compound of the present invention are also included in thepresent invention. The salt can be produced in accordance with aconventional method using the compound of a formula (I) or (I′) providedby the present invention.

Specifically, when the compound of the formula (I) or (I′) has, forexample, a basic group derived from an amino group, a pyridyl group andthe like in the molecule, the compound can be converted into acorresponding salt by treating with an acid.

Examples of the acid addition salt include hydrohalide salts such ashydrochloride, hydrofluoride, hydrobromide and hydroiodide; inorganicacid salts such as nitrate, perchlorate, sulfate, phosphate andcarbonate; lower alkyl sulfonic acid salts such as methanesulfonate,trifluoromethanesulfonate and ethanesulfonate; aryl sulfonic acid saltssuch as benzenesulfonate and p-toluenesulfonate; organic acid salts suchas fumarate, succinate, citrate, tartrate, oxalate and maleate; and acidaddition salts of amino acids, such as glutamate and aspartate.

Also, when the compound of the present invention has an acidic groupsuch as a carboxyl group in the molecule, the compound can also beconverted into a corresponding pharmaceutically acceptable salt bytreating with a base. Examples of the base addition salt include alkalimetal salts such as sodium and potassium; alkali earth metal salts suchas calcium and magnesium; and salts of organic bases, such as ammoniumsalts, guanidine, triethylamine and dicyclohexylamine.

Furthermore, the compound of the present invention may be present as afree compound, or arbitrary hydrate or solvate of a salt thereof.

Depending on selection of starting materials and methods, the compoundsof the present invention may exist as one form in possible isomers ormixtures thereof, for example, substantially pure geometrical (cis- ortrans-) isomers, optical isomers (enantiomers, antipodes), racemicforms, or mixtures thereof. The above-mentioned possible isomers ormixtures thereof fall within the scope of the present invention.

All obtainable isomer mixtures can be separated into pure geometrical oroptical isomers, diastereomers or racemic forms based on aphysicochemical difference of component by, for example, chromatographyand/or fractional crystallization.

All of the obtained racemic forms of the final product or intermediatecan be optically resolved into optical antipodes by a known method, forexample, a diastero isomer salt obtained from an optically active acidor base compound is separated and each optically active acid or basecompound is isolated. The products from the racemic forms can also beresolved by chiral chromatography, for example, high performance liquidchromatography using a chiral adsorbent.

In starting compounds and precursors which are converted into thecompounds of the present invention by the method in the presentdescription, existing functional groups such as amino, thiol, carboxyland hydroxy groups may be optionally protected with a commonconventional protecting group in preparative organic chemistry. The thusprotected amino, thiol, carboxyl and hydroxy groups can be convertedinto free amino, thiol, carboxyl and hydroxy groups under mildconditions without causing breakage of a molecular framework or theother undesirable minor reaction.

The protecting group is inserted so as to protect the functional groupfrom an undesirable reaction with a reaction component under theconditions used to perform a desired chemical conversion. Necessity andselection of the protecting group for a specific reaction are known tothose skilled in the art, and depend on properties of the functionalgroup to be protected (hydroxy group, amino group, etc.), structure andstability of the molecule with the substituent constituting a partthereof, and reaction conditions. Examples of the protecting groupinclude OTs, OTHP, methoxymethyl and OAc. The protecting group ispreferably a protecting group which is eliminated under acidicconditions.

In the diagnosis of tauopathy, the compound of the present invention canbe used as a probe without labeling. For example, the presence orabsence of the portion to be stained may be examined by bringing thecompound of the present invention into contact with a biopsy tissuesample. However, it is common to use the labeled compound of the presentinvention as a probe for the diagnosis of tauopathy. Examples of labelinclude a fluorescent substance, an affinity substance, an enzymesubstrate, a radioactive nuclide and the like. A probe labeled with aradioactive nuclide is usually used in image diagnosis of tauopathy. Itis possible to label the compound of the present invention with variousradioactive nuclides by the methods which are well known in the art. Forexample, ³H, ¹⁴C, ³⁵S, ¹³¹I and the like are radioactive nuclides whichhave been used for a long time, and is often utilized in vivo. Generalrequirements for imaging diagnosis probes and means for their detectionare to allow making an in vivo diagnosis, to cause less harm to patients(particularly, to be non-invasive), to have a high sensitivity ofdetection, to have an appropriate half-life (to have an appropriateperiod of time for preparing the labeled probes and for diagnosis) andthe like. Accordingly, it has recently tended to employ positronemission tomography (PET) utilizing γ-ray displaying a high sensitivityand permeability of materials or computed tomography (SPECT) with γ-rayemitting nuclides. Among them, PET, which detects two γ-rays emitting inopposite directions from a positron emitting nuclide by means ofsimultaneous counting with a pair of detectors, provides informationwhich is excellent in resolution and quantification and thus ispreferable. For SPECT, the compound of the present invention can belabeled with a γ-ray emitting nuclide such as ^(99m)Tc, ¹¹¹In, ⁶⁷Ga,²⁰¹Tl, ¹²³I, ¹³³Xe and the like. ^(99m)Tc and ¹²³I are often used forSPECT. For PET, the compound of the present invention can be labeledwith a positron emitting nuclide such as ¹¹C, ¹³N, ¹⁵O, ¹⁸F, ⁶²Cu, ⁶⁴Cu,⁶⁸Ga, ⁷⁶Br and the like. Among positron emitting nuclides, ¹¹C, ¹³N, ¹⁵Oand ¹⁸F are preferable, ¹⁸F and ¹¹C are more preferable, ¹⁸F isparticularly preferable, from the viewpoint of having an appropriatehalf-life, the ease of labeling and the like. Although the position oflabeling the compound of the present invention with a radiation emittingnuclide such as a positron emitting nuclides or γ-ray emitting nuclidecan be any position, labeling may preferably be carried out at an alkylgroup and on phenyl ring in the compound. Such labeled compounds of thepresent invention are also included in the present invention. Forexample, when the compound of the present invention is labeled with ¹⁸F,any position of the side chain may be labeled with ¹⁸F, or hydrogen onthe ring may be substituted with ¹⁸F. For example, hydrogen contained inanyone of alkyl substituents may be substituted with ¹⁸F. Also, when thecompound of the present invention is labeled with ¹¹C, carbon containedin any one of alkyl substituents in the side chain may be substitutedwith ¹¹C. Although it is obvious to a person with an ordinary skill inthe art, m of ^(99m)Tc denotes a nuclear isomer in a quasi-stable state.

Radionuclides used in the compounds according to the present inventionare generated on an instrument termed cyclotron or generator. A personwith an ordinary skill in the art can select methods and instruments forproduction depending upon nuclides to be produced. Nuclides thusproduced can be used to label the compounds of the present invention.

Methods of producing labeled compounds, which have been labeled withthese radionuclides, are well known in the art. Typical methods includechemical synthesis, isotope exchange, and biosynthesis processes.Chemical synthesis processes have been traditionally and widelyemployed, and are essentially the same as usual chemical synthesisprocesses, except that radioactive starting materials are used. Variousnuclides are introduced into compounds by these chemical processes.Isotope exchanging processes are processes by which ³H, ³⁵S, ¹²⁵I or thelike contained in a compound of a simple structure is transferred intocompound having a complex structure, thereby obtaining a compound havinga complex structure that has been labeled with these nuclide.Biosynthesis processes are processes by which a compound labeled with¹⁴C, ³⁵S or the like is given to cells such as microorganisms to obtainits metabolites having these nuclide introduced therein.

With respect to the labeling position, similarly to usual synthesis,synthetic schemes can be designed, depending upon the purpose, so that alabel can be introduced at a desired position. Such design is well knownto a person with ordinary skill in the art.

When utilizing positron emitting nuclides such as ¹¹C, ¹³N, ¹⁵O and ¹⁸F,which have relatively short half-lives, for example, it is also possibleto generate a desired nuclide from a (highly) small-sized cyclotronplaced in a facility such as hospital, which in turn is used to label adesired compound at its desired position by any of the above-describedmethods, followed by carrying out immediately diagnosis, examination,treatment or the like.

These methods well known to a person with ordinary skill in art, andenable one to carry out labeling by introducing a desired nuclide intothe compound of the present invention at its desired position.

The compound of the present invention, which has been labeled, may beadministered to subjects locally or systemically. Routes foradministration include intradermal, intraperitoneal, intravenous,intra-arterial injections or infusions into the spinal fluid and thelike, and can be selected depending on factors such as the disease type,nuclide used, compound used, the condition of the subject, the site tobe examined. The site to be examined can be investigated with means suchas PET, SPECT by administering the probe of the present invention,followed by the elapse of a sufficient time to allow its binding to tauprotein and decay. These procedures can be selected as appropriatedepending on factors such as the disease type, nuclide used, compoundused, the condition of the subject, the site to be examined.

The dose of the compound of the present invention, which has beenlabeled with a radionuclide, varies depending on the disease type,nuclide used, compound used, the age, physical condition, and gender ofthe subject, the degree of the disease, the site to be examined and thelike. In particular, sufficient care has to be taken about exposuredoses to the subject. For example, the amount of radioactivity of thecompound labeled with a positron emitting nuclide such as ¹¹C, ¹³N, ¹⁵Oand ¹⁸F of the present invention, is usually within a range from 3.7megabecquerels to 3.7 gigabecquerels, and preferably from 18megabecquerels to 740 megabecquerels.

The compound of the present invention or a salt or solvate thereof issuited for use in a treatment method of tauopathy, a diagnosis method, acomposition for treatment, a composition for diagnosis, a kit fordiagnosis, use for the production of these compositions and kits, andother uses, which will be described below. The compounds or salts orsolvates thereof exemplified in the above description about thecompounds of formulae (I) to (VI) are preferable, and those included inthe compound of formula (I) or a salt or solvate thereof areparticularly preferable. Among the compounds of the present invention,compounds having, as R², R³ or R⁶ in formula (I),

are suited for administration to the human body because of considerablyless or scarce accumulation in bone.

The present invention provides a composition for image diagnosis oftauopathy, containing the compound of the present invention. Thecomposition of the present invention contains the compound of thepresent invention and a pharmaceutically acceptable carrier. It ispreferred that the compound of the present invention in the compositionis labeled. Although various labeling methods are possible as describedabove, labeling with radionuclides (in particular, positron emittingnuclides such as ¹¹C, ¹³N, ¹⁵O and ¹⁸F for PET) is desirable for in vivoimage diagnosis applications. It is preferable from their purposes thatthe form of the composition of the present invention is one allowinginjection or infusion. Accordingly, a pharmaceutically acceptablecarrier is preferably liquid and examples thereof include, but are notlimited to, aqueous solvents such as potassium phosphate buffer,physiological saline, ringer solution and distilled water; andnon-aqueous solvents such as polyethylene glycol, vegetable oil,ethanol, glycerin, dimethylsulfoxide and propylene glycol. A mixingratio of the carrier to the compound of the present invention can beappropriately selected depending on the site of application, detectionmeans and the like, and is usually from 100,000:1 to 2:1, and preferablyfrom 10,000:1 to 10:1. The composition of the present invention mayfurther contain known antimicrobials (for example, antimicrobial drug,etc.), local anesthetics (for example, procaine hydrochloride, etc.),buffers (for example, Tris-hydrochloride buffer, HEPES buffer, etc.),osmolytes (for example, glucose, sorbitol, sodium chloride, etc.) andthe like.

Furthermore, the present invention provides a kit for image diagnosis oftauopathy, containing the compound of the present invention as theessential ingredient. Usually, the kit is a package in which each of thecomponents such as the compound of the present invention, a solvent fordissolving the compound, a buffer, an osmoregulatory agent, anantimicrobial, a local anesthetic are packaged separately intorespective containers, or some of the components are packaged togetherinto respective containers. The compound of the present invention may beunlabeled or labeled. When not labeled, the compound of the presentinvention can be labeled, prior to use, by usual methods as describedabove. In addition, the compound of the present invention may bepresented as a solid, such as a lyophilized powder, or in solution inappropriate solvents. Solvents may be similar to carriers used in theabove composition of the present invention. Each of the components suchas a buffer, an osmoregulatory agent, an antimicrobial, a localanesthetic, also may be similar to those used in the above compositionof the present invention. While various containers can be selected asappropriate, they may be of shapes suitable for carrying out theintroduction of a label into the compound of the present invention, orof light-shielding materials, depending on the nature of compounds, ortake forms such as vials or syringes, so as to be convenient foradministration to patients. The kit may also contains, as appropriate,tools necessary for diagnosis, for example, syringes, an infusion set,or device for use in a PET or SPECT apparatus. The kit usually has itsinstructions attached thereto.

Furthermore, the compounds of the present invention are specificallybound to tau protein, and thus the compounds of the present inventioncan be also used, for example, for detecting and quantifying tau proteinwith or without labeling by contacting with sample specimens in vitro.For example, the compounds of the present invention can be used forstaining tau protein in microscopic specimens, for colorimetricdetermination of tau protein in samples, or for quantifying tau proteinusing a scintillation counter. Preparation of a microscope specimen andstaining using the compound of the present invention can be carried outby a conventional method known to a person with an ordinary skill in theart.

As described above, the compounds of the present invention are highlyspecific to tau protein. Therefore, the compounds of the presentinvention are useful, for example, for studies of disease with tauprotein accumulation or in their diagnosis before and after death, andcould be useful, for example, as agents for staining neurofibrillarytangles in brain sections of Alzheimer's disease patients. Staining ofspecimens, for example, brain sections using the compounds of thepresent invention can be carried out in a conventional method known to aperson of ordinary skill in the art.

As described above, among the compounds of the present invention,compounds having, as R², R³ or R⁶ in formula (I):

may cause considerably less or scarce accumulation in bone. Accordingly,these compounds of the present invention are not only considerably safeprobes for the diagnosis of tauopathy, but also exhibit high safety evenwhen used as remedies or preventives described hereinafter.

Accordingly, the present invention is directed to a composition forstaining of amyloid β protein, particularly tau in a sample, containingthe compound of the present invention or a pharmaceutically acceptablesalt or solvate thereof, and a kit for staining of amyloid β protein,particularly tau in a sample, containing the compound of the presentinvention or a pharmaceutically acceptable salt or solvate thereof asessential ingredients. Furthermore, the present invention is directed toa method of staining amyloid β protein, particularly tau in a sample,the method comprising using the compound of the present invention or apharmaceutically acceptable salt or solvate thereof. Samples suited forabove staining are brain sections.

As described above, it has been found that neurotoxicity is present inamyloid β protein or tau of a β-sheet structure. It is considered thatthe compound of the present invention is specifically bound to amyloid βprotein of a β-sheet structure, particularly tau, and thus neurotoxicityis inhibited. Accordingly, it is considered that the compound of thepresent invention serves as remedies or preventives for causes of adisease, particularly tauopathy, for example, Alzheimer's disease sinceprotein itself has a β-sheet structure.

Accordingly, the present invention provides:

a method of treating and/or preventing diseases with amyloid β proteinaccumulation, particularly tauopathy, the method comprisingadministering a compound of the formula (I) or a salt or solvatethereof;

a method of diagnosing diseases with amyloid β protein accumulation,particularly tauopathy, the method comprising using a compound of theformula (I) or a salt or solvate thereof; and

use of a compound of the formula (I) or a salt or solvate thereof forthe production of a composition or kit for the treatment, prevention ordiagnosis of diseases with amyloid β protein accumulation, particularlytauopathy.

Forms of such pharmaceutical compositions are not limited in particular,but liquid formulations, particularly formulations for injection, arepreferable. Such formulations for injection can be infused directly intothe brain, or alternatively the above pharmaceutical compositions can beformulated for intravenous injection or drip and administered, since thecompounds of the present invention have high permeability through theblood-brain barrier, as shown in the Examples. Such liquid formulationscan be prepared by methods well known in the art. Solutions can beprepared, for example, by dissolving the compound of the presentinvention in an appropriate carrier, water for injection, phisilogicalsaline, Ringer's solution or the like, sterilizing the solution througha filter or the like, and then filling the sterilized solution intoappropriate containers, for example, vials or ampules. Solutions alsocan be lyophilized and when used, reconstituted with an appropriatecarrier. Suspensions can be prepared, for example, by sterilizing thecompound of the present invention, for example, by exposure to ethyleneoxide, and then suspending it in a sterilized liquid carrier.

When such a pharmaceutical composition is used in a liquid formulation,particularly a formulation for injection, an injection can be preparedby adding a solubilizing agent to a quinoline derivative according tothe present invention.

It is possible to use, as the solubilizing agent, nonionic surfactants,cationic surfactants, amphoteric surfactants and the like used in theart. Among these solubilizing agents, Polysorbate 80, polyethyleneglycol, ethanol or propylene glycol is preferable, and Polysorbate 80 ismore preferable.

The amount of the compounds of the present invention to be administeredto a human subject in the above treatment method, prevention method anduse varies depending on the condition, gender, age, weight of thepatient and the like, and is generally within a range from 0.1 mg to 1g, preferably from 1 mg to 100 mg, and more preferably from 5 mg to 50mg, per day for adult humans weighing 70 kg. It is possible to conduct atreatment with such a dose for a specified period of time, followed byincreasing or reducing the dose according to the outcome.

Furthermore, the compound of the present invention or a pharmaceuticallyacceptable salt or solvate thereof can also be used as a probe for thediagnosis of conformational disease, particularly tauopathy, preferablyan image diagnosis probe labeled with a radiation nuclide. Furthermore,the compounds of the present invention have the effect for the treatmentand/or prevention of conformational disease, particularly tauopathy.

Accordingly, the present invention is also directed to:

a compound of the present invention used as an image diagnosis probe ofconformational disease, particularly tauopathy, or a salt or solvatethereof;

a composition or kit for image diagnosis of conformational disease,particularly tauopathy, comprising the compound of the present inventionor a salt or solvate thereof;

a pharmaceutical composition for the prevention and/or treatment ofconformational disease, particularly tauopathy, comprising a compound ofthe present invention or a pharmaceutically acceptable salt or solvatethereof, and a pharmaceutically acceptable carrier;

a method of diagnosing conformational disease, particularly tauopathy,the method comprising using a compound of the present invention or apharmaceutically acceptable salt or solvate thereof;

use of a compound of the present invention or a pharmaceuticallyacceptable salt or solvate thereof for the diagnosis of conformationaldisease, particularly tauopathy;

a method of preventing and/or treating conformational disease,particularly tauopathy, the method comprising administering a compoundof the present invention or a pharmaceutically acceptable salt orsolvate thereof to the subject;

use of a compound of the present invention or a pharmaceuticallyacceptable salt or solvate thereof for the prevention and/or treatmentof conformational disease, particularly tauopathy; and

use of a compound of the present invention in the production of apharmaceutical composition for the prevention and/or treatment ofconformational disease, particularly tauopathy.

The dose of the compounds of the present invention to be administered toa human subject in the above treatment methods and prevention methods isas described above.

The compounds of the present invention recognize neurofibrillary tanglescontaining excessively phosphorylated tau protein as a constituentingredient, and thus the compounds can be used as a probe for thedetection of neurofibrillary tangles, or as an agent for stainingneurofibrillary tangles. Accordingly, the present invention also relatesto use of the compound of the present invention or a salt or solvatethereof as a probe for detecting neurofibrillary tangles, particularly aprobe for the diagnoasis of images. Examples of compounds of the presentinvention which are preferable for staining of neurofibrillary tanglesinclude THK-5004, THK-5035, THK-5038, THK-5051, THK-5058, THK-5064,THK-5065, THK-5066, THK-5071, THK-5077, THK-5078, THK-5105, THK-5106,THK-5107, THK-5112, THK-5116, THK-5117, THK-5122 and the like.

Accordingly, the present invention provides a composition for detectingor staining neurofibrillary tangles, containing the compound of thepresent invention or a salt or solvate thereof;

a kit for detecting or staining neurofibrillary tangles, containing thecompound of the present invention or a salt or solvate thereof;

a method of detecting or staining neurofibrillary tangles, whichcomprises using the compound of the present invention or a salt orsolvate thereof; and

use of the compound of the present invention or a salt or solvatethereof for the production of a composition for detecting or stainingneurofibrillary tangles.

Methods known in a person with an ordinary skill in the art can beapplied to methods of preparing and staining a sample specimen in thedetection or staining of the above neurofibrillary tangles.

Further, the present invention provides a kit for preparing a compoundof the present invention or a pharmaceutically acceptable salt orsolvate thereof, the kit comprising a compound of the present inventionor a pharmaceutically acceptable salt or solvate thereof, a labelingagent, and optionally instructions for labeling the compound. Thelabeling agent is, for example, a radioactive nuclide or a positronemitting nuclide. The radioactive nuclide is, for example, a γ-rayemitting nuclide. The positron emitting nuclide is selected from, forexample, the group consisting of ¹¹C, ¹³N, ¹⁵O, ¹⁸F, ³⁵MCl, ⁷⁶Br, ⁴⁵Ti,⁴⁸V, ⁶⁰Cu, ⁶¹Cu, ⁶²Cu, ⁶⁴Cu, ⁶⁶Ga, ⁸⁹Zr, ^(94m)Tc, and ¹²⁴I. Preferably,the positron emitting nuclide is ¹¹C or ¹⁸F. The labeling agent is anagent that is suitable for labeling the compound, and is known to thoseskilled in the art.

Typical examples of the compound of the formula (I) used preferably inthe present invention are shown below.

TABLE 1-1 THK-5004

2-(4-aminophenyl)-8-(1- fluoromethyl-2-hydroxyethoxy) quinoline THK-5035

2-(4-diethylaminophenyl)- 6-(1-fluoromethyl-2- hydroxy)quinolineTHK-5038

2-(4-diethylaminophenyl)- 7-(2-fluoromethyl-2- hydroxyethoxy)quinolineTHK-5051

2-(4-diethylaminophenyl)- 8-(1-fluoromethyl-2- hydroxyethoxy)quinoline

TABLE 1-2 THK-5058

2-(4-diethylaminophenyl)- 7-(1-fluoromethyl-2- hydroxyethoxy)quinolineTHK-5059

2-(4-diethylaminophenyl)- 4-(3-fluoro-2-hydroxypropoxy) quinolineTHK-5064

2-(4-diethylaminophenyl)- 5-(1-fluoromethyl-2- hydroxyethoxy)quinolineTHK-5065

2-(4-diethylaminophenyl)- 3-(1-fluoromethyl-2- hydroxyethoxy)quinoline

TABLE 1-3 THK-5066

2-(4-diethylaminophenyl)-8- [(3-fluoro-2-hydroxy)propoxy] quinolineTHK-5071

2-(4-fluoromethyl-2-hydroxyethoxy)- 2-(4-dimethylaminophenyl) quinolineTHK-5077

7-(1-fluoromethyl-2-hydroxyethoxy)- 2-(4-methylaminophenyl) quinolineTHK-5078

2-(4-ethylmethylaminophenyl)- 7-(1-fluoromethyl-2-hydroxyethoxy)-quinoline THK-5105

6-[(3-fluoro-2-hydroxy)propoxy]- 2-(4-dimethylaminophenyl) quinoline

TABLE 1-4 THK-5106

7-[(3-fluoro-2-hydroxy) propoxy]-2-(4- methylaminophenyl) quinolineTHK5107

7-[(3-fluoro-2-hydroxy) propoxy]-2-(4- dimethylaminophenyl) quinolineTHK-5112

2-(4-ethylmethylaminophenyl)- 7-[(3-fluoro-2-hydroxy) propoxy]quinolineTHK-5116

2-(4-aminophenyl)-6-[(3- fluoro-2-hydroxy)propoxy] quinoline THK-5117

6-[(3-fluoro-2-hydroxy) propoxy]-2-(4- methylaminophenyl) quinoline

TABLE 1-5 THK-5122

6-[(3-fluoro-2-hydroxy) propoxy]-2-(4- diethylaminophenyl) quinolineTHK-5075

7-amino-2-(4-fluorophenyl) quinoline THK-5076

2-(4-fluorophenyl)-7- dimethylaminoquinoline THK-5079

5-amino-2-(4-fluorophenyl) quinoline THK-5080

2-(4-fluorophenyl)-5- dimethylaminoquinoline oxalate

TABLE 1-6 THK-5081

8-amino-2-(4-fluorophenyl) quinoline THK-5082

2-(4-fluorophenyl)-8- dimethylaminoquinoline THK-5086

6-amino-2-(4-fluorophenyl) quinoline THK-5087

2-(4-fluorophenyl)-6- dimethylaminoquinoline THK-932 

2-(2-aminopyrid-5-yl)-7- (1-fluoromethyl-2- hydroxyethoxy)quinoline

TABLE 1-7 THK-5100

6-ethylmethylamino-2-(4- fluorophenyl)quinoline THK-5088

6-diethylamino-2-(2- fluoropyrid-5-yl)quinoline THK-5089

8-ethylmethylamino-2-(2- fluoropyrid-5-yl) quinoline THK-5091

5-ethylamino-2-(2- fluoropyrid-5-yl)quinoline

TABLE 1-8 THK-5092

5-diethylamino-2-(2- fluoropyrid-5-yl)quinoline THK-5097

7-diethylamino-2-(2- fluoropyrid-5-yl)quinoline THK-5098

7-ethylmethylamino-2-(2- fluoropyrid-5-yl)quinoline THK-5125

2-(4-ethylaminophenyl)-6- [(3-fluoro-2-hydroxy) propoxy]quinoline

TABLE 1-9 THK-5127

2-(2-aminopyrid-5-yl)- 6-[(3-fluoro-2-hydroxy) propoxy]quinolineTHK-5151

2-(2-methylaminopyrid-5- yl)-6-[(3-fluoro-2-hydroxy) propoxy]quinolineTHK-5129

6-[(3-fluoro-2-hydroxy) propoxy]-2-(2-dimethyl- aminopyrid-5-yl)quinoline THK-5130

2-(2-diethylaminopyrid- 5-yl)-6-[(3-fluoro- 2-hydroxy)propoxy] quinolineTHK-5142

2-(2-ethylaminopyrid- 5-yl)-6-[(3-fluoro-2- hydroxy)propoxy] quinoline

TABLE 1-10 THK-5177

1-fluoro-3-{2-[4-(4-methyl piperazin-1-yl)phenyl]quinolin-6-yloxy)propan-2-ol THK-5178

1-fluoro-3-{2-[6-(piperazin-1- yl)pyridin-3-yl]quinolin-6-yloxy}propan-2-ol THK-5180

1-fluoro-3-{2-[6-(4-methyl piperazin-1-yl)pyridin-3-yl]quinolin-6-yloxy} propan-2-ol

TABLE 1-11 THK-5136

6-[(3-fluoro-2-hydroxy) propoxy]-2-(4-methyl-3,4-dihydro-2H-pyrido[3,2-b] [1,4]oxazin-7-yl) quinoline THK-5153

6-[(3-fluoro-2-hydroxy) propoxy]-2-(1-methyl-1,2,3,4-tetrahydroquinolin- 6-yl)quinoline THK-5157

6-[(3-fluoro-2-hydroxy-1,1- dimethyl)propoxy]-2-(1-methyl-1,2,3,4-tetrahydro- quinolin-6-yl)quinoline

TABLE 1-12 THK-5128

2-(4-amino-3-fluorophenyl)- 6-dimethylaminoquinoline THK-5147

2-[4-(amino)-3-[(3-fluoro- 2-hydroxy)propoxy]phenyl]-6-methylaminoquinoline THK-5148

2-[3-(3-fluoro-2-hydroxy-1,1- dimethyl)propoxy]-4-(dimethylamino)-phenyl]-6- dimethylaminoquinoline THK-5155

6-amino-2-[4-(amino)-3-[(3- fluoro-2-hydroxy)propoxy] phenyl]quinoline

TABLE 1-13 THK-5156

2-[3-[(3-fluoro-2-hydroxy) propoxy]-4-(dimethyl- amino)phenyl]-6-dimethylaminoquinoline THK-5158

2-[3-[(3-fluoro-2-hydroxy) propoxy]-4-(methylamino)-phenyl]-6-methylamino- quinoline THK-5159

2-[4-(amino)-3-[(3-fluoro- 2-hydroxy)propoxy] phenyl]-6-dimethylaminoquinoline THK-5160

6-amino-2-[3-[(3-fluoro- 2-hydroxy)propoxy]-4- (dimethylamino)phenyl]-quinoline THK-5161

2-[3-[(3-fluoro-2-hydroxy) propoxy]-4-(dimethyl- amino)phenyl]-6-methylaminoquinoline

TABLE 1-14 THK-5162

2-[3-[2-[2-(2-fluoro ethoxy)ethoxy]ethoxy- 4-(methylamino)phenyl]-6-dimethylaminoquinoline THK-5164

2-[3-[(3-fluoro-2- hydroxy)propoxy]-4- (methylamino)phenyl]-6-dimethylaminoquinoline THK-5165

6-amino-2-[3-[(3-fluoro- 2-hydroxy)propoxy]-4- (methylamino)phenyl]-quinoline

TABLE 1-15 THK-5154

2-[3-[(3-fluoro-2- hydroxy)propoxy]-2- (dimethylamino)pyrid-5-yl]-6-dimethylamino- quinoline THK-5166

2-[3-[(3-fluoro-2- hydroxy)propoxy]-2- (dimethylamino)pyrid-5-yl]quinoline

TABLE 1-16 THK-5170

6-[(3-fluoro-2-hydroxy) propoxy]-2-(6-fluoro- pyridin-3-yl)quinolineTHK-5171

6-[(3-fluoro-2-hydroxy) propoxy]-2-(4- methoxyphenyl) quinoline THK-5172

6-[(3-fluoro-2-hydroxy) propoxy]-2-[4- (hydroxymethyl)phenyl] quinolineTHK-5173

6-[(3-fluoro-2-hydroxy) propoxy]-2-(4- ethanonephenyl) quinolineTHK-5174

6-[(3-fluoro-2-hydroxy) propoxy]-2-(6- methoxypyridin-3-yl) quinoline

TABLE 1-17 THK-5175

6-[(3-fluoro-2-hydroxy) propoxy]-2-(4- ethoxyphenyl) quinoline THK-5176

6-[(3-fluoro-2-hydroxy) propoxy]-2-(4- amino-3-methoxyphenyl) quinolineTHK-5179

6-[(3-fluoro-2-hydroxy) propoxy]-2-(benzamido- 4-yl)quinoline THK-5181

6-[(3-fluoro-2-hydroxy) propoxy]-2-(3- aminophenyl)quinoline THK-5182

6-[(3-fluoro-2-hydroxy) propoxy]-2-(1- methyl-pyrazol-4-yl) quinoline

Typical examples of the compound of formula (I′) used preferably in thepresent invention are shown below. Among these compounds, THK-5039 canbe used as a synthetic precursor of THK-5035, THK-5041 can be used as asynthetic precursor of THK-5004, THK-5050 can be used as a syntheticprecursor of THK-5051, THK-5070 can be used as a synthetic precursor ofTHK-5066, THK-5072 can be used as a synthetic precursor of THK-5058,THK-5073 can be used as a synthetic precursor of THK-5038, THK-5090 canbe used as a synthetic precursor of THK-5071, THK-5095 can be used as asynthetic precursor of THK-5077, THK-5096 can be used as a syntheticprecursor of THK-5078, THK-5099 can be used as a synthetic precursor ofTHK-5064, THK-5111 can be used as a synthetic precursor of THK-5112,THK-5113 can be used as a synthetic precursor of THK-5106, THK-5115 canbe used as a synthetic precursor of THK-5107, THK-5119 can be used as asynthetic precursor of THK-5117, THK-5120 can be used as a syntheticprecursor of THK-5122, THK-5121 can be used as a synthetic precursor ofTHK-5105, THK-5123 can be used as a synthetic precursor of THK-5116,THK-5131 can be used as a synthetic precursor of THK-5125, THK-5150 canbe used as a synthetic precursor of THK-5127, THK-5152 can be used as asynthetic precursor of THK-5151, THK-5135 can be used as a syntheticprecursor of THK-5129, THK-5138 can be used as a synthetic precursor ofTHK-5130, THK-5143 can be used as a synthetic precursor of THK-5142,THK-5163 can be used as a synthetic precursor of THK-5164, THK-5167 canbe used as a synthetic precursor of THK-5136, and THK-5168 can be usedas a synthetic precursor of THK-5156.

TABLE 2-1 THK-5039

2-(4-diethylaminophenyl)- 6-[(2-hydroxy-1-tosyloxymethyl)ethoxy]quinoline THK-5041

2-(4-aminophenyl)-8-(2-hydroxy- 1-tosyloxymethylethoxy) quinoline

TABLE 2-2 THK-5050

2-(4-diethylaminophenyl)- 8-(2-hydroxy-1-tosyloxy methylethoxy)quinolineTHK-5070

2-(4-diethylaminophenyl)- 8-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy] propoxy]quinoline THK-5072

2-(4-diethylaminophenyl)- 7-(2-hydroxy-1-tosyloxy methylethoxy)quinolineTHK-5073

2-(4-diethylaminophenyl)- 7-[[(2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy] propoxy]quinoline THK-5090

7-(2-hydroxy-1-tosyloxy- methylethoxy)-2-(4- dimethylaminophenyl)quinoline THK-5095

7-(2-hydroxy-1-tosyloxy- methylethoxy)-2-(4-methyl aminophenyl)quinolineTHK-5096

2-(4-ethylmethylamino- phenyl)-7-(2-hydroxy-1- tosyloxymethylethoxy)quinoline

TABLE 2-3 THK-5099

2-(4-diethylaminophenyl)- 5-(2-hydroxy-1-tosyloxy methylethoxy)quinolineTHK-5111

2-(4-ethylmethylaminophenyl)- 7-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy] propoxy]quinoline THK-5113

2-(4-methylaminophenyl)- 7-[[2-(tetrahydro-2H- pyran-2-yloxy)-3-tosyloxy]propoxy]quinoline THK-5115

2-(4-dimethylaminophenyl)- 7-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy] quinoline THK-5119

2-(4-methylaminophenyl)- 6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy] quinoline THK-5120

2-(4-diethylaminophenyl)- 6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy] quinoline

TABLE 2-4 THK-5121

2-(4-dimethylaminophenyl)- 6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy] quinoline THK-5123

2-(4-aminophenyl)-6-[[2- (tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy] quinoline

TABLE 2-5 THK-5131

2-(4-ethylaminophenyl)- 6-[[2-(tetrahydro-2H- pyran-2-yloxy)-3-tosyloxy]propoxy] quinoline THK-5150

2-(2-aminopyrid-5-yl)- 6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy]propoxy] quinoline THK-5152

2-(2-methylaminopyrid- 5-yl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy] propoxy]quinoline THK-5135

2-(2-dimethylaminopyrid- 5-yl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy] propoxy]quinoline THK-5138

2-(2-diethylaminopyrid- 5-yl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy] propoxy]quinoline THK-5143

2-(2-ethylaminopyrid- 5-yl)-6-[[2-(tetrahydro-2H-pyran-2-yloxy)-3-tosyloxy] propoxy]quinoline

TABLE 2-6 THK-5163

2-[4-(methylamino)- 3-[[2-(tetrahydro-2H- pyran-2-yloxy)-3-tosyloxy]propoxy] phenyl]-6-dimethyl- aminoquinoline THK-5167

6-[[2-(tetrahydro-2H- pyran-2-yloxy)-tosyloxy] propoxy]-2-(4-methyl-3,4-dihydro-2H-pyrido [3,2-b][1,4]oxazin-7-yl) quinoline THK-5168

2-[4-(dimethylamino)- 3-[[2-(tetrahydro-2H- pyran-2-yloxy)-3-tosyloxy]propoxy] phenyl]-6-dimethyl- aminoquinoline

The present invention will be descried in more detail and specificallybelow by way of Examples, but the present invention is not limited tothe Examples.

EXAMPLE 1 Synthesis of the Compounds of the Present Invention

Silica gel BW300 manufactured by Fuji Silysia Chemical Ltd. was used insilica gel column chromatography of Examples. Chromatorex NH-DM1020manufactured by Fuji Silysia Chemical Ltd. was used in basic silica gelcolumn chromatography using an amino group-bonded type silica gel.

¹H-NMR was measured using UNITY INOVA500 (500 MHz) manufactured byVARIAN, JNM-LA400 (400 MHz) manufactured by JEOL, Ltd. and Gemini 2000(300 MHz) tetramethylsilane manufactured by VARIAN as standardsubstances, and all δ values were measured by ppm.

Mass spectrum was measured by atmospheric pressure chemical ionization(APCI) using LCQ-Advantage manufactured by ThermoQuest or SSQ-7000Cmanufactured by FinniganMAT.

Infrared spectra were measured using a Paragon1000 FT-IR manufactured byPerkin-Elmer, and B-545 manufactured by BUCHI was used for themeasurement of a melting point.

Meanings of abbreviations in the measurement of NMR are shown below.

s: singlet

d: doublet

dd: double doublet

ddd: double double doublet

t: triplet

dt: double triplet

q: quartet

m: multiplet

br: broad

J: coupling constant

Hz: hertz

CDCl₃: deuterated chloroform

DMSO-d₆: deuterated dimethylsulfoxide

Synthesis Examples and Label Example of the compounds of the presentinvention are shown below.

Synthesis Method of THK-5004

Synthesis of 3

To a mixture of 1 (1.29 g, 8.0 mmol), 2 (1.66 g, 9.0 mmol),triphenylphosphine (2.36 g, 9.0 mmol) and tetrahydrofuran (20 ml), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (1.82g, 9.0 mmol) was added dropwise under ice cooling and stirring, and themixture was stirred at room temperature for 16 hours. The solvent of thereaction solution was distilled off under reduced pressure and theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=2/3) to obtain 3 (2.62 g, 100%) as apale yellow oily substance.

Synthesis of 5

To a methylene chloride (20 ml) solution of 3 (2.62 g, 8.0 mmol) and 4(2.2 g, 10.4 mmol), a methylene chloride (10 ml) solution oftrifluoromethanesulfonic anhydride (2.7 g, 9.6 mmol) was added dropwiseunder ice cooling and stirring, and the mixed solution was stirred atroom temperature for 16 hours. 4 (2.2 g, 10.4 mmol) andtrifluoromethanesulfonic anhydride (2.7 g, 9.6 mmol) were added,followed by further stirring at room temperature for 16 hours. To thereaction solution, ice water was added and the solution was extractedwith chloroform. The extraction liquid was dried and the solvent wasdistilled off under reduced pressure, and then the residue was purifiedby silica gel column chromatography (eluting solvent: n-hexane/ethylacetate=20/1) to obtain 5 (1.47 g, 40%) as a colorless oily substance.

APCI-MS m/z 460[M+H]⁺

Synthesis of 7

To a mixture of 5 (1.45 g, 3.15 mmol), 6a (1.00 g, 3.15 mmol) and1,2-dimethoxyethane (20 ml), an aqueous 2M sodium carbonate solution(3.5 ml, 7.0 mmol) and tetrakistriphenylphosphine palladium (146 mg,0.126 mmol) were added and the mixture was heated at reflux for 16 hoursunder an argon atmosphere. The reaction solution was allowed to returnto room temperature and the solvent was distilled off under reducedpressure, and then water was added to the residue and the solution wasextracted with chloroform. The extraction liquid was dried and thesolvent was distilled off under reduced pressure, and then the residuewas purified by silica gel column chromatography (eluting solvent:n-hexane/ethyl acetate=9/1) to obtain 7 (1.55 g, 98%) as a pale yellowfoam-like substance.

APCI-MS m/z 503[M+H]⁺

Synthesis of THK-5004

To a mixture of 7 (1.54 g, 3.0 mmol) and anisole (0.98 ml),methanesulfonic acid (4.9 ml) was added and the mixture was stirred atroom temperature for 16 hours. To the reaction solution, ice water wasadded and the solution was made basic with an aqueous potassiumcarbonate solution and then extracted with chloroform. The extractionliquid was dried and the solvent was distilled off under reducedpressure, the residue was purified by silica gel column chromatography(eluting solvent: n-hexane/ethyl acetate=2/1→3/2→1/4) and then washedwith diisopropylether to obtain THK-5004 (593 mg, 62%) as a yellowsolid.

mp 97-100° C., ¹H NMR (400 MHz, CDCl₃) δ 3.70-3.80 (2H, m), 3.80-3.90(2H, brs), 4.40-4.50 (1H, m), 4.70 to 5.00 (2H, m), 6.29 (1H, t, J=6.4Hz), 6.75-6.85 (2H, m), 7.49 (1H, t, J=7.6 Hz), 7.59 (1H, d, J=7.6 Hz),7.82 (1H, d, J=8.8 Hz), 7.92 (2H, d, J=8.8 Hz), 8.19 (1H, d, J=8.8 Hz)

IR (Nujol) 1600 cm⁻¹

APCI-MS m/z 313[M+H]⁺

Synthesis Method of THK-5035

Synthesis of 9

To an N,N-dimethylformamide (50 ml) suspension of 8 (3.00 g, 18.6 mmol),potassium carbonate (2.83 g, 20.5 mmol) and benzyl chloride (2.25 ml,19.6 mmol) were added and the mixture was stirred at 105° C. for 1 hour.The reaction solution was allowed to return to room temperature andwater was added, and then the solution was extracted with warm ethylacetate. The extraction liquid was washed with water and dried, and thensolvent was concentrated to about 100 ml under reduced pressure. Afterbeing allowed to cool, the precipitated crystal was collected byfiltration and then dried to obtain 9 (2.66 g, 57%) as colorlesscrystals.

mp 220-221° C., IR (Nujol) 1661, 1623 cm⁻¹

APCI-MS m/z 252[M+H]⁺

Synthesis of 10

To a methylene chloride (70 ml) suspension of 9 (2.66 g, 10.6 mmol) and4 (3.04 g, 14.8 mmol), trifluoromethanesulfonic anhydride (2.14 ml, 12.7mmol) was added dropwise under ice cooling and stirring, and the mixturewas stirred at room temperature for 2 hours. 4 (1.52 g, 7.41 mmol) andtrifluoromethanesulfonic anhydride (1.07 ml, 6.35 mmol) were added,followed by further stirring at room temperature for 3 days. Water wasadded to the reaction solution and the solution was extracted with ethylacetate. The extraction liquid was washed with water and dried, and thenthe solvent was distilled off under reduced pressure. The residue waspurified by silica gel column chromatography (eluting solvent:n-hexane/ethyl acetate=20/1, 9/1) and then recrystallized from ethylacetate-n-hexane to obtain 10 (3.75 g, 92%) as a colorless crystal.

mp 109.5-110° C., APCI-MS m/z 384[M+H]⁺

Synthesis of 11

To a mixture of 10 (2.19 g, 5.7 mmol), 6 (1.65 g, 6.0 mmol) and1,2-dimethoxyethane (20 ml), an aqueous 2M sodium carbonate solution(5.5 ml, 11 mmol) and tetrakistriphenylphosphine palladium (277 mg,0.240 mmol) were added and the mixture was stirred under an argonatmosphere at 90° C. for 16 hours. The reaction solution was allowed toreturn to room temperature and insolubles were removed by filtration andthen washed with chloroform. The filtrate and the wash were combined andthe mixture was extracted with chloroform. The extraction liquid waswashed with saturated saline and then dried, and the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: chloroform) to obtain 11(1.86 g, 85%) as an orange solid.

APCI-MS m/z 383[M+H]⁺

Synthesis of 12

To a mixture of 11 (1.85 g, 4.84 mmol) and anisole (1 ml),methanesulfonic acid (5 ml) was added and the mixture was stirred atroom temperature for 30 minutes. To the reaction solution, ice water wasadded and the solution was washed with ethyl acetate. The aqueous layerwas made basic with sodium hydrogen carbonate and extracted withchloroform-tetrahydrofuran. The extraction liquid was dried and thesolvent was distilled off under reduced pressure, and then the residuewas washed with n-hexane to obtain 12 (1.25 g, 88%) as an orange solid.

APCI-MS m/z 293[M+H]⁺

Synthesis of 13

To a mixture of 12 (468 mg, 1.6 mmol), 2 (354 mg, 1.9 mmol),triphenylphosphine (504 mg, 1.9 mmol) and tetrahydrofuran (20 ml), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (388mg, 1.9 mmol) was added dropwise under ice cooling and stirring, and themixture was stirred at room temperature for 16 hours. Triphenylphosphine(150 mg, 0.57 mmol) and diisopropyl azodicarboxylate (120 mg, 0.59 mmol)were added, followed by further stirring at room temperature for 2hours. The solvent of the reaction solution was distilled off underreduced pressure and the residue was purified by silica gel columnchromatography (eluting solvent: chloroform) to obtain 13 (733 mg, 100%)as an orange solid.

Synthesis of 14

To a mixture of 13 (733 mg, 1.6 mmol) and anisole (1 ml),methanesulfonic acid (5 ml) was added under ice cooling and stirring,and the mixture was stirred at room temperature for 50 minutes. To thereaction solution, ice water (40 ml) was added and the solution waswashed with ethyl acetate. The aqueous layer was made basic withpotassium carbonate and extracted with chloroform. The extraction liquidwas dried and the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: chloroform/methanol=99/1) and then washed with diisopropyletherto obtain 14 (400 mg, 68%) as a yellow solid.

APCI-MS m/z 369[M+H]⁺

Synthesis of THK-5035

To an ethanol (10 ml) solution of 14 (400 mg, 1.09 mmol), 4M HCl/ethylacetate (1 ml, 4 mmol) was added and the solvent was distilled off underreduced pressure. The residue was washed with diethyl ether and thenrecrystallized from isopropanol-acetone to obtain THK-5035 (368 mg, 84%)as orange crystals.

mp 207-209° C., ¹H NMR (400 MHz, DMSO-d₆+D₂O) δ 2.53 (6H, t, J=7.0 Hz),3.52 (4H, q, J=7.0 Hz), 4.6-4.9 (3H, m), 6.99 (2H, d, J=8.5 Hz), 7.7-7.8(2H, m), 8.13 (2H, d, J=9.1 Hz), 8.27 (2H, d, J=9.1 Hz), 8.71 (1H, d,J=9.1 Hz)

IR (Nujol) 1595, 1460, 1216 cm⁻¹

APCI-MS m/z 369[M+H]⁺

Synthesis Method of THK-5038

Synthesis of 17

To a mixture of 15 (1.00 g, 6.21 mmol), 16 (1.55 g, 7.45 mmol),triphenylphosphine (1.95 g, 7.45 mmol) and tetrahydrofuran (80 ml),diisopropyl azodicarboxylate (1.48 ml, 7.45 mmol) was added dropwiseunder ice cooling and stirring, and the mixture was stirred at roomtemperature for 16 hours. The solvent of the reaction solution wasdistilled off under reduced pressure and the residue was purified bysilica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/2) to obtain 17 (1.80 g, 82%) as a pale pink solid.

mp 132-135° C., APCI-MS m/z 352[M+H]⁺

Synthesis of 18

To a methylene chloride (25 ml) solution of 17 (1.80 g, 5.12 mmol) and 4(1.47 g, 7.17 mmol), trifluoromethanesulfonic anhydride (1.03 ml, 6.14mmol) was added dropwise under ice cooling and stirring, and the mixturewas stirred at room temperature for 70 minutes. Water was added to thereaction solution and the solution was extracted with ethyl acetate. Theextraction liquid was washed with water and dried, and then the solventwas distilled off under reduced pressure. The residue was purified bysilica gel column chromatography (eluting solvent:n-hexane→n-hexane/ethyl acetate=50/1→30/1) to obtain 18 (2.09 g, 84%) asa pale pink oily substance.

APCI-MS m/z 484[M+H]⁺

Synthesis of 19

To a mixture of 18 (1.20 g, 2.48 mmol), 6 (680 mg, 2.48 mmol) and1,2-dimethoxyethane (30 ml), potassium carbonate (1.03 g, 7.44 mmol) andwater (0.62 ml) were added, and tetrakistriphenylphosphine palladium(124 mg, 0.124 mmol) was added under an argon atmosphere, followed bystirring at 80° C. for 2.5 hours. The reaction solution was allowed toreturn to room temperature, water was added and the solution wasextracted with ethyl acetate. The extraction liquid was washed withwater and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: n-hexane/ethyl acetate=20/1) and then recrystallizedfrom n-hexane to obtain 19 (700 mg, 59%) as pale yellow crystals.

mp 102-102.5° C., APCI-MS m/z 483[M+H]⁺

Synthesis of THK-5038

To a tetrahydrofuran (10 ml) solution of 19 (700 mg, 1.45 mmol), 1Mtetra-n-butylammonium fluoride/tetrahydrofuran (1.45 ml, 1.45 mmol) wasadded and the mixture was stirred at room temperature for 1 hour. Waterwas added to the reaction solution and the solution was extracted withethyl acetate. The extraction liquid was washed with water and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: n-hexane/ethyl acetate=2/1) and then recrystallized from ethylacetate-n-hexane to obtain THK-5038 (482 mg, 90%).

mp 110-110.5° C., ¹H NMR (500 MHz, DMSO-d₆) δ 1.14 (6H, t, J=7.0 Hz),3.42 (4H, q, J=7.0 Hz), 4.10-4.19 (3H, m), 4.45 to 4.64 (2H, m), 5.55(1H, d, J=4.8 Hz), 6.78 (2H, d, J=9.0 Hz), 7.12 (1H, dd, J=8.7, 2.6 Hz),7.35 (1H, d, J=2.6 Hz), 7.81 (1H, d, J=9.0 Hz), 7.85 (1H, d, J=9.0 Hz),8.10 (2H, d, J=9.0 Hz), 8.21 (1H, d, J=9.0 Hz)

IR (Nujol) 1622, 1596 cm⁻¹

APCI-MS m/z 369[M+H]⁺

Synthesis Method of THK-5039

Synthesis of 21

To a mixture of 12 (400 mg, 1.37 mmol), 20 (592 mg, 1.64 mmol),triphenylphosphine (431 mg, 1.64 mmol) and tetrahydrofuran (20 ml),diisopropyl azodicarboxylate (0.325 ml, 1.64 mmol) was added dropwiseunder ice cooling and stirring, and the mixture was stirred at roomtemperature for 16 hours. 20 (592 mg, 1.64 mmol), triphenylphosphine(431 mg, 1.64 mmol) and tetrahydrofuran (10 ml) were added, followed byfurther stirring at room temperature for 24 hours. The solvent of thereaction solution was distilled off under reduced pressure and theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/9) to obtain 21 (870 mg, 100%) as ayellow oily substance.

APCI-MS m/z 635[M+H]⁺

Synthesis of THK-5039

To a chloroform (12 ml) solution of 21 (870 mg, 1.37 mmol),trifluoroacetic acid (8 ml) and then water (2 ml) were added dropwiseunder ice cooling and stirring, and the mixture was stirred at roomtemperature for 1.5 hours. To the reaction solution, ice water and ethylacetate were added, and the solution was extracted with ethyl acetateafter adjusting the pH to 9 using an aqueous potassium carbonatesolution. The extraction liquid was washed with water and dried, andthen the solvent was distilled off under reduced pressure. The residuewas purified by silica gel column chromatography (eluting solvent:n-hexane/ethyl acetate=4/1, 2/1) and then recrystallized from ethylacetate-n-hexane to obtain THK-5039 (219 mg, 31%).

mp 119-120° C., ¹1-1NMR (500 MHz, DMSO-d₆) δ 1.14 (6H, t, J=7.0 Hz),2.33 (3H, s), 3.42 (4H, q, J=7.0 Hz), 4.07 (5H, m), 5.62 (1H, d, J=4.8Hz), 6.78 (2H, d, J=9.0 Hz), 7.21 (2H, m), 7.40 (2H, d, J=8.0 Hz), 7.78(2H, d, J=8.0 Hz), 7.86 (1H, d, J=8.0 Hz), 7.97 (1H, d, J=8.7 Hz), 8.07(2H, d, J=8.7 Hz), 8.20 (1H, d, J=8.7 Hz)

APCI-MS m/z 521[M+H]⁺

Synthesis Method of THK-5050

Synthesis of 23

To a mixture of 1 (2.00 g, 12.4 mmol), 22 (4.06 g, 14.9 mmol),triphenylphosphine (3.91 g, 14.9 mmol) and tetrahydrofuran (40 ml),diisopropyl azodicarboxylate (2.95 ml, 14.9 mmol) was added dropwiseunder ice cooling and stirring, and the mixture was stirred at roomtemperature for 3 days. The solvent of the reaction solution wasdistilled off under reduced pressure and the residue was purified bysilica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/4, 2/3) to obtain 23 (3.45 g, 67%) as a pale yellowoily substance.

APCI-MS m/z 416[M+H]⁺

Synthesis of 24

To a methylene chloride (40 ml) solution of 23 (3.44 g, 8.28 mmol) and 4(2.38 g, 11.6 mmol), trifluoromethanesulfonic anhydride (1.67 ml, 9.94mmol) was added dropwise under ice cooling and stirring, and the mixturewas stirred at room temperature for 1 hour. Water was added to thereaction solution and the solution was extracted with ethyl acetate. Theextraction liquid was washed with water and dried, and then the solventwas distilled off under reduced pressure. The residue was purified bysilica gel column chromatography (eluting solvent:n-hexane→n-hexane/ethyl acetate=9/1) to obtain 24 (4.16 g, 92%) as acolorless oily substance.

APCI-MS m/z 548[M+H]⁺

Synthesis of 25

To a mixture of 24 (2.00 g, 3.65 mmol), 6 (1.01 g, 3.65 mmol) and1,2-dimethoxyethane (30 ml), potassium carbonate (1.51 g, 11 mmol) andwater (0.63 ml) were added, and tetrakistriphenylphosphine palladium(210 mg, 0.183 mmol) was added under an argon atmosphere and the mixturewas stirred at 80° C. for 2 hours. The reaction solution was allowed toreturn to room temperature and water was added, and then solution wasextracted with ethyl acetate. The extraction liquid was washed withwater and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: n-hexane/ethyl acetate=9/1) to obtain 25 (1.95 g, 98%)as a pale yellow oily substance.

APCI-MS m/z 547[M+H]⁺

Synthesis of 26

To a mixture of 25 (1.94 g, 3.55 mmol) and anisole (3 ml),methanesulfonic acid (9 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 20minutes. To the reaction solution, ice water and then ethyl acetate wereadded, and the solution was extracted with ethyl acetate after adjustingthe pH to 9 using an aqueous potassium carbonate solution. Theextraction liquid was washed with saturated saline and dried, and thenthe solvent was distilled off under reduced pressure. The residue waspurified by silica gel column chromatography (eluting solvent:n-hexane/ethyl acetate=4/1→1/2→1/3) to obtain 26 (1.22 g, 94%) as ayellow powder.

APCI-MS m/z 367[M+H]⁺

Synthesis of 27

To a 1,2-dimethoxyethane (120 ml) solution of 26 (1.36 g, 3.71 mmol),paratoluenesulfonic anhydride (1.21 g, 3.71 mmol) and triethylamine(0.78 ml, 5.57 mmol) were added and the mixture was stirred at roomtemperature for 16 hours. The reaction solution was washed withsaturated saline and dried, and then the solvent was distilled off underreduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: n-hexane/ethyl acetate=3/1) to obtain27 (500 mg, 26%) as an orange foam-like substance.

Synthesis of THK-5050

27 (638 mg, 1.23 mmol) was dissolved in ethyl acetate and the solutionwas subjected to short silica gel column chromatography (elutingsolvent: ethyl acetate) to remove an origin substance. After dissolvingby adding oxalic acid (221 mg, 2.45 mmol) to the eluate, the solvent wasconcentrated to about 100 ml under reduced pressure. After dissolvingthe precipitate by adding ethanol (100 ml), the solvent was concentratedto about 100 ml under reduced pressure. Insolubles were removed byfiltration and the filtrate was concentrated to about 50 ml underreduced pressure. Isopropanol (50 ml) was added and the solvent wasconcentrated under reduced pressure to precipitate a crystal. Afterstanding at 5° C. for 16 hours, the precipitated crystals were collectedby filtration and dried to obtain THK-5050 (520 mg, 69%) as orangecrystals.

mp 124-125° C., ¹H NMR (400 MHz, DMSO-d₆) δ 1.15 (6H, t, J=7.0 Hz), 2.30(3H, s), 3.50 (4H, q, J=7.0 Hz), 3.82-3.88 (2H, m), 4.42 to 4.49 (1H,m), 4.85 (1H, dd, J=14, 3.0 Hz), 4.92 (1H, dd, J=14, 9.4 Hz), 6.93 (2H,d, J=9.1 Hz), 7.15 (2H, d, J=7.9 Hz), 7.46 (2H, d, J=7.9 Hz), 7.66-7.70(3H, m), 7.85 (1H, t, J=8.2 Hz), 7.93 (1H, dd, J=8.2, 1.2 Hz), 8.06 (1H,d, J=9.0 Hz), 8.95 (1H, d, J=9.0 Hz)

IR (Nujol) 1633, 1590 cm⁻¹

APCI-MS m/z 521[M+H]⁺

Synthesis Method of THK-5051

Synthesis of 29

To a mixture of 28 (894 mg, 2.30 mmol), 6 (706 mg, 2.57 mmol) and1,2-dimethoxyethane (10 ml), an aqueous 2M sodium carbonate solution(2.5 ml, 5.0 mmol) and tetrakistriphenylphosphine palladium (119 mg,0.10 mmol) were added under an argon atmosphere and the mixture wasstirred at 90° C. for 2 hours. The reaction solution was allowed toreturn to room temperature and insolubles were removed by filtration,and then the solution was washed with chloroform. The filtrate and thewash were combined and the mixture was extracted with chloroform. Theextraction liquid was dried and the solvent was distilled off underreduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: n-hexane/ethyl acetate=9/1→4/1) toobtain 29 (880 mg, 100%) as a pale yellow oily substance.

APCI-MS m/z 383 [M+H]⁺

Synthesis of 30

To a mixture of 29 (880 mg, 2.30 mmol) and anisole (1 ml),methanesulfonic acid (5 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 1 hour. Tothe reaction solution, ice water was added and the solution was washedwith ethyl acetate. The aqueous layer was extracted with chloroformafter adjusting the pH to 9 using an aqueous saturated sodium hydrogencarbonate solution. The extraction liquid was dried and the solvent wasdistilled off under reduced pressure to obtain 30 (670 mg, 100%) as ayellow oily substance.

APCI-MS m/z 293[M+H]⁺

Synthesis of 31

To a mixture of 30 (670 mg, 2.3 mmol), 2 (516 mg, 2.8 mmol),triphenylphosphine (918 mg, 3.5 mmol) and tetrahydrofuran (10 ml), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (708mg, 3.5 mmol) was added dropwise under ice cooling and stirring, and themixture was stirred at room temperature for 16 hours. The solvent of thereaction solution was distilled off under reduced pressure and theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/19→1/9) to obtain 31 (780 mg, 73%) asa pale green oily substance.

APCI-MS m/z 459[M+H]⁺

Synthesis of 32

To a mixture of 31 (775 mg, 1.69 mmol) and anisole (1 ml),methanesulfonic acid (5 ml) was added under ice cooling and stirring,and the mixture was stirred at room temperature for 1.5 hours. To thereaction solution, ice water was added and the solution was washed withethyl acetate. The aqueous layer was made basic with an aqueoussaturated sodium hydrogen carbonate solution and extracted withchloroform. The extraction liquid was dried and the solvent wasdistilled off under reduced pressure, and then the residue was purifiedby silica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/4) to obtain 32 (608 mg, 97%) as an orange oilysubstance.

APCI-MS m/z 369[M+H]⁺

Synthesis of THK-5051

To an acetone solution of 32 (576 mg, 1.56 mmol), oxalic acid (281 mg,3.12 mmol) was added to form an oxalate, which was recrystallized frommethanol-diethyl ether to obtain THK-5051 (414 mg, 53%) as orangecrystals.

mp 126-128° C., ¹H NMR (400 MHz, DMSO-d₆+D₂O) δ 1.15 (6H, t, J=7.0 Hz),3.44 (4H, q, J=7.0 Hz), 3.6-3.8 (1H, m), 4.7 to 4.9 (3H, m), 6.84 (2H,d, J=9.0 Hz), 7.39 (1H, d, J=7.4 Hz), 7.44 (1H, t, J=7.7 Hz), 7.60 (1H,d, J=7.7 Hz), 8.05 (1H, d, J=9.0 Hz), 8.12 (2H, d, J=9.0 Hz), 8.35 (1H,d, J=9.0 Hz)

APCI-MS m/z 369[M+H]⁺

Synthesis Method of THK-5058

Synthesis of 33

To a mixture of 15 (1.00 g, 6.2 mmol), 2 (1.37 g, 7.45 mmol),triphenylphosphine (1.95 g, 7.45 mmol) and tetrahydrofuran (50 ml),diisopropyl azodicarboxylate (1.48 ml, 7.45 mmol) was added dropwiseunder ice cooling and stirring, and the mixture was stirred at roomtemperature for 16 hours. To the reaction solution, 2 (0.67 g, 3.73mmol), triphenylphosphine (0.98 g, 3.73 mmol), tetrahydrofuran (10 ml)and diisopropyl azodicarboxylate (0.74 ml, 3.73 mmol) were added,followed by further stirring at room temperature for 6 hours. Thesolvent of the reaction solution was distilled off under reducedpressure and the residue was purified by silica gel columnchromatography (eluting solvent: chloroform,chloroform/methanol/concentrated ammonia water=50/1/0.1) to obtain 33(1.66 g, 82%) as a pale brown oily substance.

APCI-MS m/z 328[M+H]⁺

Synthesis of 34

To a methylene chloride (25 ml) solution of 33 (1.65 g, 5.0 mmol) and 4(1.45 g, 7.06 mmol), trifluoromethanesulfonic anhydride (1.02 ml, 6.05mmol) was added dropwise under ice cooling and stirring, and the mixturewas stirred at room temperature for 2 hours. 4 (0.21 g, 1.01 mmol) andtrifluoromethanesulfonic anhydride (0.17 ml, 1.01 mmol) were added,followed by further stirring at room temperature for 30 minutes. To thereaction solution, ice water was added and the solution was extractedwith ethyl acetate. The extraction liquid was washed with water anddried, and then the solvent was distilled off under reduced pressure.The residue was purified by silica gel column chromatography (elutingsolvent: n-hexane, n-hexane/ethyl acetate=4/1) to obtain 34 (1.94 g,84%) as a colorless solid.

mp 81-82° C., APCI-MS m/z 460[M+H]⁺

Synthesis of 35

To a mixture of 34 (1.00 g, 2.18 mmol), 6 (0.6 g, 2.18 mmol) and1,2-dimethoxyethane (18 ml), potassium carbonate (0.90 g, 6.54 mmol),water (0.38 ml) and tetrakistriphenylphosphine palladium (130 mg, 0.109mmol) were added under an argon atmosphere and the mixture was stirredat 80° C. for 1.5 hours. The reaction solution was allowed to return toroom temperature and water was added, and the solution was extractedwith ethyl acetate. The extraction liquid was washed with water anddried, and then the solvent was distilled off under reduced pressure.The residue was purified by silica gel column chromatography (elutingsolvent: n-hexane/ethyl acetate=9/1, 6/1) to obtain 35 (1.00 g, 100%) asa pale yellow oily substance.

APCI-MS m/z 459[M+H]⁺

Synthesis of THK-5058

To a mixture of 35 (0.99 g, 2.16 mmol) and anisole (3 ml),methanesulfonic acid (9 ml) was added under ice cooling and stirring,and the mixture was stirred at room temperature for 1 hour. To thereaction solution, ice water and then ethyl acetate were added. Thesolution was made basic with an aqueous potassium carbonate solution andextracted with ethyl acetate. The extraction liquid was dried and thesolvent was distilled off under reduced pressure, and then the residuewas purified by silica gel column chromatography (eluting solvent:n-hexane/ethyl acetate=4/1, 2/1) and then washed with n-hexane/ethylacetate (4/1) to obtain THK-5058 (636 mg, 80%) as pale yellow crystals.

mp 89-90° C., ¹H NMR (500 MHz, DMSO-d₆) δ 1.14 (6H, t, J=7.0 Hz), 3.42(4H, q, J=7.0 Hz), 3.65-3.77 (2H, m), 4.65-4.87 (3H, m), 5.14 (1H, t,J=5.6 Hz), 6.78 (2H, d, J=8.7 Hz), 7.17 (1H, dd, J=8.8, 2.4 Hz), 7.47(1H, d, J=2.4 Hz), 7.82 (1H, d, J=8.8 Hz), 7.85 (1H, d, J=8.8 Hz), 8.10(2H, d, J=8.8 Hz), 8.21 (1H, d, J=8.8 Hz)

IR (Nujol) 1598 cm⁻¹

APCI-MS m/z 369[M+H]⁺

Synthesis Method of THK-5059

Synthesis of 37

To a mixture of 36 (1.68 g, 4.38 mmol), 6 (1.21 g, 4.38 mmol) and1,2-dimethoxyethane (36 ml), potassium carbonate (1.82 g, 13.2 mmol),water (0.76 ml) and tetrakistriphenylphosphine palladium (250 mg, 0.22mmol) were added under an argon atmosphere and the mixture was stirredat 80° C. for 1.5 hours. The reaction solution was allowed to return toroom temperature and water was added, and the solution was extractedwith ethyl acetate. The extraction liquid was washed with water anddried, and then the solvent was distilled off under reduced pressure.The residue was purified by silica gel column chromatography (elutingsolvent: n-hexane/ethyl acetate=20/1, 9/1) to obtain 37 (1.73 g) as apale yellow oily substance.

APCI-MS m/z 383[M+H]⁺

Synthesis of 38

To a mixture of 37 (1.73 g) and anisole (4 ml), methanesulfonic acid (12ml) was added under ice cooling and stirring, and the mixture wasstirred at room temperature for 3 days. To the reaction solution, icewater and then ethyl acetate were added. The solution was made basicwith an aqueous potassium carbonate solution and extracted with ethylacetate. The extraction liquid was dried and the solvent was distilledoff under reduced pressure, and then the residue was washed withn-hexane/ethyl acetate (1/1) and then dried to obtain 38 (1.20 g, 94%from 36) as pale yellow crystals.

mp 290-291° C., APCI-MS m/z 293[M+H]⁺

Synthesis of 39

To a mixture of 38 (600 mg, 2.05 mmol), 2 (454 mg, 2.46 mmol),triphenylphosphine (646 mg, 2.46 mmol) and tetrahydrofuran (20 ml),diisopropyl azodicarboxylate (0.49 ml, 2.46 mmol) was added dropwiseunder ice cooling and stirring, and the mixture was stirred at roomtemperature for 16 hours. The solvent of the reaction solution wasdistilled off under reduced pressure and the residue was purified bysilica gel column chromatography (eluting solvent: n-hexane/ethylacetate=9/1) to obtain 39 (940 mg, 100%) as a pale yellow oilysubstance.

APCI-MS m/z 459[M+H]⁺

Synthesis of 40

To a mixture of 39 (940 mg, 2.05 mmol) and anisole (2 ml),methanesulfonic acid (6 ml) was added under ice cooling and stirring,and the mixture was stirred at room temperature for 10 minutes. To thereaction solution, ice water and then ethyl acetate were added. Thesolution was made basic with an aqueous potassium carbonate solution andextracted with ethyl acetate. The extraction liquid was dried and thesolvent was distilled off under reduced pressure, and then the residuewas purified by silica gel column chromatography (eluting solvent:n-hexane/ethyl acetate=4/1, 2/1) and then recrystallized from ethylacetate to obtain 40 (627 mg, 83%) as pale yellow crystals.

mp 146-147° C., APCI-MS m/z 369[M+H]⁺

Synthesis of THK-5059

To an ethyl acetate (100 ml) solution of 40 (700 mg, 1.90 mmol), anethyl acetate suspension (10 g/20 ml) of silica gel was added and themixture was stirred at room temperature for 3 days. The silica gel wasremoved by filtration and the solution was washed with ethyl acetate.The filtrate and the wash were combined and the solvent was distilledoff under reduced pressure. The residue was recrystallized fromn-hexane/ethyl acetate (1/1) to obtain THK-5059 (484 mg, 69%) as paleyellow crystals.

mp 155-155.5° C., ¹H NMR (400 MHz, DMSO-d₆) δ 1.15 (6H, t, J=7.0 Hz),3.42 (4H, q, J=7.0 Hz), 4.19-4.41 (3H, m), 4.61 (1H, ddd, J=48, 9.7, 5.0Hz), 4.65 (1H, ddd, J=48, 10, 4.5 Hz), 5.64 (1H, d, J=5.4 Hz), 6.77 (2H,d, J=9.1 Hz), 7.42 (1H, s), 7.42-7.47 (1H, m), 7.66-7.71 (1H, m), 7.89(1H, d, J=8.2 Hz), 8.10-8.13 3H, m)

IR (Nujol) 3150, 1610 cm⁻¹

APCI-MS m/z 369[M+H]⁺

Synthesis Method of THK-5064

Synthesis of 42

To a mixture of 41 (520 mg, 3.23 mmol), 2 (710 mg, 3.87 mmol),triphenylphosphine (1.02 g, 3.87 mmol) and tetrahydrofuran (30 ml),diisopropyl azodicarboxylate (0.77 ml, 3.87 mmol) was added dropwiseunder ice cooling and stirring, and the mixture was stirred at roomtemperature for 16 hours. To the reaction solution, 2 (360 mg, 1.94mmol), triphenylphosphine (0.51 g, 1.94 mmol), diisopropylazodicarboxylate (0.38 ml, 1.94 mmol) and tetrahydrofuran (10 ml) wereadded, followed by further stirring at room temperature for 2 hours. Thesolvent of the reaction solution was distilled off under reducedpressure and the residue was purified by silica gel columnchromatography (eluting solvent: n-hexane/ethyl acetate=2/1, 1/1) toobtain 42 (600 mg, 57%) as a pale brown solid.

APCI-MS m/z 328[M+H]⁺

Synthesis of 43

To a methylene chloride (15 ml) solution of 42 (590 mg, 1.80 mmol) and 4(520 mg, 2.52 mmol), trifluoromethanesulfonic anhydride (0.36 ml, 2.16mmol) was added dropwise under ice cooling and stirring, and the mixturewas stirred at room temperature for 1 hour. 4 (74 mg, 0.36 mmol) andtrifluoromethanesulfonic anhydride (0.061 ml, 0.37 mmol) were added,followed by further stirring at room temperature for 30 minutes. To thereaction solution, ice water was added and the solution was extractedwith ethyl acetate. The extraction liquid was washed with water anddried, and then the solvent was distilled off under reduced pressure.The residue was purified by silica gel column chromatography (elutingsolvent: n-hexane, n-hexane/ethyl acetate=9/1) to obtain 43 (560 mg,68%) as a pale yellow oily substance.

APCI-MS m/z 460[M+H]⁺

Synthesis of 44

To a mixture of 43 (550 mg, 1.20 mmol), 6 (330 mg, 1.20 mmol) and1,2-dimethoxyethane (20 ml), potassium carbonate (500 mg, 3.59 mmol),water (0.21 ml) and tetrakistriphenylphosphine palladium (69 mg, 0.06mmol) were added under an argon atmosphere, and the mixture was stirredat 80° C. for 1.5 hours. The reaction solution was allowed to return toroom temperature, diluted with ethyl acetate and washed with water. Theorganic layer was dried and the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: n-hexane/ethyl acetate=4/1) to obtain 44 (550 mg,100%) as a pale yellow oily substance.

APCI-MS m/z 459[M+H]⁺

Synthesis of THK-5064

To a mixture of 44 (540 mg, 1.18 mmol) and anisole (1.0 ml),methanesulfonic acid (3.0 ml) was added under ice cooling and stirring,and the mixture was stirred at room temperature for 20 minutes. To thereaction solution, ice water and then ethyl acetate were added. Thesolution was made basic with an aqueous potassium carbonate solution andextracted with ethyl acetate. The extraction liquid was washed withwater and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: n-hexane/ethyl acetate=4/1) to obtain THK-5064 (403mg, 93%) as an orange powder.

¹H NMR (500 MHz, DMSO-d₆) δ 1.14 (6H, t, J=7.0 Hz), 3.42 (4H, q, J=7.0Hz), 3.73-3.78 (2H, m), 4.70-4.88 (3H, m), 5.14 (1H, t, J=5.3 Hz), 6.79(2H, d, J=9.0 Hz), 7.10 (1H, d, J=7.4 Hz), 7.56 (1H, d, J=8.7 Hz), 7.61(1H, t, J=8.4 Hz), 7.98 (1H, d, J=8.7 Hz), 8.11 (2H, d, J=9.0 Hz), 8.51(1H, d, J=9.0 Hz)

IR (Nujol) 3400, 1610 cm⁻¹

APCI-MS m/z 369[M+H]⁺

Synthesis Method of THK-5065

Synthesis of 46

To a mixture of 45 (3.0 g, 18.6 mmol), 2 (3.5 g, 19.0 mmol),triphenylphosphine (5.77 g, 22.0 mmol) and tetrahydrofuran (60 ml),diisopropyl azodicarboxylate (4.45 g, 22.0 mmol) was added dropwiseunder ice cooling and stirring, and the mixture was stirred at roomtemperature for 16 hours. The solvent of the reaction solution wasdistilled off under reduced pressure and the residue was purified bysilica gel column chromatography (eluting solvent: n-hexane/ethylacetate=2/1, 1/1) to obtain 46 (3.0 g, 49%) as a colorless solid.

mp 124-125° C.

Synthesis of 47

To a methylene chloride (20 ml) solution of 46 (2.62 g, 8.0 mmol) and 4(2.2 g, 10.4 mmol), a methylene chloride (10 ml) solution oftrifluoromethanesulfonic anhydride (2.7 g, 9.6 mmol) was added dropwiseunder ice cooling and stirring, and the mixture was stirred at roomtemperature for 16 hours. 4 (2.2 g, 10.4 mmol) andtrifluoromethanesulfonic anhydride (2.7 g, 9.6 mmol) were added,followed by further stirring at room temperature for 16 hours. To thereaction solution, ice water was added and the solution was extractedwith chloroform. The extraction liquid was dried and the solvent wasdistilled off under reduced pressure, and then the residue was purifiedby silica gel column chromatography (eluting solvent: n-hexane/ethylacetate=20/1) to obtain 47 (1.47 g, 40%) as a colorless oily substance.

APCI-MS m/z 460[M+H]⁺

Synthesis of 48

To a mixture of 47 (1.20 g, 2.61 mmol), 6 (0.72 g, 2.61 mmol) and1,2-dimethoxyethane (30 ml), sodium carbonate (560 mg, 5.28 mmol), water(3 ml) and tetrakistriphenylphosphine palladium (150 mg, 0.13 mmol) wereadded under an argon atmosphere, and the mixture was heated at refluxfor 3 hours. The reaction solution was allowed to return to roomtemperature, diluted with ethyl acetate, washed in turn with water andsaturated saline and then dried. The solvent was distilled off underreduced pressure and the residue was purified by silica gel flash columnchromatography (eluting solvent: n-hexane/ethyl acetate=8/1, 6/1) toobtain 48 (1.06 g, 88%) as a pale yellow oily substance.

APCI-MS m/z 459[M+H]⁺

Synthesis of THK-5065

To a mixture of 48 (1.27 g, 2.77 mmol) and anisole (1.0 ml),methanesulfonic acid (10 ml) was added and the mixture was stirred atroom temperature for 30 minutes. The reaction solution was diluted withethyl acetate and poured into ice water, followed by liquid separation.The aqueous layer was made basic with concentrated ammonia water andextracted with ethyl acetate. The extraction liquid was washed in turnwith water and saturated saline and then dried. The solvent wasdistilled off under reduced pressure and the residue was recrystallizedfrom diisopropylether to obtain THK-5065 (758 mg, 74%) as pale yellowcrystals.

mp 135-136° C., ¹H NMR (400 MHz, CDCl₃) δ 1.20 (6H, t, J=7.1 Hz), 1.95(1H, br), 3.42 (4H, q, J=7.1 Hz), 3.77-3.88 (2H, m), 4.56-4.65 (1H, m),4.65-4.69 (1H, m), 4.77-4.81 (1H, m), 6.75 (2H, d, J=9.0 Hz), 7.47 (1H,m), 7.57 (1H, m), 7.61 (1H, s), 7.68 (1H, dd, J=8.2, 1.5 Hz), 7.93 (2H,d, J=9.0 Hz), 8.07 (1H, d, J=8.5 Hz)

IR (Nujol) 3200, 1617, 1606 cm⁻¹

APCI-MS m/z 369[M+H]⁺

Synthesis Method of THK-5066

Synthesis of 49

To a mixture of 30 (648 mg, 2.2 mmol), 16 (559 mg, 2.7 mmol),triphenylphosphine (880 mg, 3.36 mmol) and tetrahydrofuran (10 ml), atetrahydrofuran (5 ml) solution of diisopropyl azodicarboxylate (678 mg,3.36 mmol) was added dropwise, and the mixture was stirred at roomtemperature for 16 hours. 16 (447 mg, 2.15 mmol), triphenylphosphine(700 mg, 2.67 mmol) and diisopropyl azodicarboxylate (542 mg, 2.68 mmol)were added, followed by further stirring at room temperature for 16hours. The solvent of the reaction solution was distilled off underreduced pressure and the residue was purified by silica gel flash columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/9) to obtainan oily product (1.24 g). To the obtained oily product, tetrahydrofuran(8 ml) and tetrabutylammonium fluoride (2.2 ml/1M tetrahydrofuransolution) were added, followed by stirring at room temperature for 1hour. The solvent of the reaction solution was distilled off underreduced pressure and the residue was dissolved in chloroform. Thesolution was washed with an aqueous saturated sodium hydrogen carbonatesolution and dried. The solvent of the organic layer was distilled offunder reduced pressure and the residue was purified by silica gel flashcolumn chromatography (eluting solvent: ethylacetate/n-hexane=1/19→1/9→1/4→1/2) to obtain 49 (497 mg, 61%) as a paleyellow foam-like substance.

APCI-MS m/z 369[M+H]⁺

Synthesis of THK-5066

To an acetone (10 ml) solution of 49 (494 mg, 1.30 mmol), oxalic acid(181 mg, 2.01 mmol) was added to form an oxalate. Acetone was distilledoff under reduced pressure and the residue was crystallized from diethylether to obtain THK-5066 (584 mg, 87%) as a dark orange solid.

¹H NMR (500 MHz, DMSO-d₆) δ 1.15(6H, q, J=7.0 Hz), 3.43 (4H, q, J=7.0Hz), 4.15-4.30 (3H, m), 4.6-4.8 (2H, m), 6.81 (2H, d, J=9.0 Hz), 7.24(1H, d, J=7.7 Hz), 7.40 (1H, t, J=7.7 Hz), 7.49 (1H, d, J=7.7 Hz), 8.03(1H, d, J=8.6 Hz), 8.14 (2H, d, J=9.0 Hz), 8.27 (1H, d, J=8.6 Hz)

IR (Nujol) 1592, 1461 cm⁻¹

APCI-MS m/z 369[M+H]⁺

Synthesis Method of THK-5070

Synthesis of 50

To a N,N-dimethylformamide (20 ml) suspension of 1 (2.00 g, 12.4 mmol),thionyl chloride (3.56 ml, 49.6 mmol) was added dropwise under icecooling and stirring, and the mixture was stirred at room temperaturefor 2 hours, and then thionyl chloride (3.56 ml, 49.6 mmol) was added,followed by further stirring at 50° C. for 3.5 hours. The reactionsolution was allowed to return to room temperature, poured into icewater and extracted with ethyl acetate. The extraction liquid was washedwith water and dried, and then the solvent was distilled off underreduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/20) to obtain50 (1.72 g, 77%) as a pale yellow solid.

mp 79-80° C., APCI-MS m/z 180/182[M+H]⁺

Synthesis of 52

To a mixture of 50 (1.71 g, 9.52 mmol), 51 (1.51 g, 11.4 mmol),triphenylphosphine (3.00 g, 11.4 mmol) and tetrahydrofuran (50 ml),diisopropyl azodicarboxylate (2.27 ml, 11.4 mmol) was added dropwiseunder ice cooling and stirring, and the mixture was stirred at roomtemperature for 16 hours. The solvent of the reaction solution wasdistilled off under reduced pressure and the residue was purified bysilica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/19, 1/9) to obtain 52 (2.05 g, 73%) as a colorlesssolid.

mp 94-95° C., APCI-MS m/z 294/296[M+H]⁺

Synthesis of 53

To a mixture of 52 (600 mg, 2.04 mmol), 6 (560 mg, 2.04 mmol) and1,2-dimethoxyethane (20 ml), potassium carbonate (850 mg, 6.12 mmol),water (0.36 ml), tetrakistriphenylphosphine palladium (120 mg, 0.102mmol) were added under an argon atmosphere, and the mixture was stirredat 80° C. for 16 hours. The reaction solution was allowed to return toroom temperature, and water and ethyl acetate were added and insolubleswere removed by filtration with celite, followed by liquid separation ofthe filtrate. The organic layer was washed with water and dried, andthen the solvent was distilled off under reduced pressure. The residuewas purified by silica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/9) to obtain 53 (820 mg, 98%) as a pale yellow solid.

mp 87-89° C.

Synthesis of 54

A mixture of 53 (820 mg, 2.02 mmol), methanol (50 ml) and 1Nhydrochloric acid (2.02 ml) was stirred at 70° C. for 1 hour. Thereaction solution was allowed to return to room temperature andneutralized with sodium hydrogen carbonate, and then ethyl acetate andsodium sulfate were added, followed by stirring at room temperature.Insolubles were removed by filtration and the solvent of the filtratewas distilled off under reduced pressure. The residue was purified bysilica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/1, ethyl acetate) to obtain 54 (720 mg, 97%) as apale yellow foam-like substance.

Synthesis of 55

To a pyridine (5 ml)-tetrahydrofuran (5 ml) solution of 54 (480 mg, 1.31mmol), a tetrahydrofuran (5 ml) solution of paratoluenesulfonicanhydride (641 mg, 1.97 mmol) was added dropwise over 20 minutes underice cooling and stirring, and the mixture was stirred at the sametemperature for 10 minutes. To the reaction solution, methanol was addedand the solvent was distilled off under reduced pressure, and thentoluene containing triethylamine was added and the solvent was distilledoff under reduced pressure. The residue was purified by silica gelcolumn chromatography (eluting solvent: ethyl acetate/n-hexane=1/2, 1/1)to obtain 55 (340 mg, 50%) as a yellow foam-like substance.

Synthesis of THK-5070

To a tetrahydrofuran (20 ml) solution of 55 (390 mg, 0.749 mmol),3,4-dihydro-2H-pyran (0.204 ml, 2.25 mmol) and paratoluenesulfonic acidmonohydrate (168 mg, 0.976 mmol) were added, and the mixture was stirredat room temperature for 18 hours, and then 3,4-dihydro-2H-pyran (0.475ml, 5.24 mmol) was added, followed by further stirring at roomtemperature for 24 hours. The reaction solution was neutralized withtriethylamine and the solvent was distilled off under reduced pressure.The residue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/2) and then purified by NH silica gelcolumn chromatography (eluting solvent: ethyl acetate/n-hexane=1/1) andcrystallized from methanol (15 ml) to obtain THK-5070 (96 mg, 21%) aspale yellow crystals.

mp 80-81° C.

IR (Nujol) 1597 cm⁻¹, APCI-MS m/z 605[M+H]⁺

Synthesis Method of THK-5071

Synthesis of 57

To a mixture of 34 (735 mg, 1.6 mmol), 56 (435 mg, 1.76 mmol) and1,2-dimethoxyethane (8 ml), an aqueous 2M sodium carbonate solution (1.6ml, 3.2 mmol) and tetrakistriphenylphosphine palladium (92 mg, 0.08mmol) were added under an argon atmosphere, and the mixture was stirredat 90° C. for 16 hours. The reaction solution was allowed to return toroom temperature and insolubles were removed by filtration, and thesolution was washed with chloroform. The filtrate and the wash werecombined and the mixture was dried, and then the solvent was distilledoff under reduced pressure. The residue was purified by silica gel flashcolumn chromatography (eluting solvent: ethyl acetate/n-hexane=1/5, 1/2)to obtain 57 (678 mg, 98%) as a pale yellow oily substance.

APCI-MS m/z 431[M+H]⁺

Synthesis of THK-5071

To a mixture of 57 (638 mg, 1.48 mmol) and anisole (5 ml),methanesulfonic acid (1 ml) was added under ice cooling and stirring,and the mixture was stirred at room temperature for 30 minutes. To thereaction solution, ice water was added and the solution was washed withethyl acetate. The aqueous layer was made basic with potassium carbonateand extracted with chloroform. The extraction liquid was dried and thesolvent was distilled off under reduced pressure, and then the residuewas purified by silica gel flash column chromatography (eluting solvent:ethyl acetate/n-hexane=1/4) and then washed withdiisopropylether-chloroform to obtain THK-5071 (394 mg, 78%) as a yellowsolid.

mp 134-135.5° C., ¹H NMR (500 MHz, DMSO-d₆) δ 3.01 (6H, s), 3.7-3.8 (2H,m), 4.7-4.9 (3H, m), 6.84 (2H, d, J=9 Hz), 7.18 (1H, dd, J=8.7, 2.3 Hz),7.48 (1H, d, J=2.3 Hz), 7.83 (1H, d, J=9 Hz), 7.88 (1H, d, J=8.7 Hz),8.13 (2H, d, J=9 Hz), 8.23 (1H, d, J=8.4 Hz)

IR (Nujol) 1597, 1505, 1202 cm⁻¹,

APCI-MS m/z 341[M+H]⁺

Synthesis Method of THK-5072

Synthesis of 58

To an N,N-dimethylformamide (30 ml) suspension of 15 (2.87 g, 17.81mmol), thionyl chloride (5.11 ml, 71.23 mmol) was added dropwise underice cooling and stirring, and the mixture was stirred at roomtemperature for 30 minutes, followed by stirring at 70° C. for 30minutes. The reaction solution was allowed to return to roomtemperature, poured into ice water and extracted with ethyl acetate. Theextraction liquid was washed with water and dried, and then the solventwas distilled off under reduced pressure. The residue was purified bysilica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/1, ethyl acetate) to obtain 58 (3.04 g, 95%) as abrownish solid.

APCI-MS m/z 180/182[M+H]⁺

Synthesis of 59

To a tetrahydrofuran (30 ml) solution of 58 (1.50 g, 8.35 mmol) andethyl vinyl ether (1.21 g, 16.7 mmol), a paratoluenesulfonic acidpyridine salt (210 mg, 0.84 mmol) was added, and the mixture was stirredat room temperature for 16 hours. After the pH of the reaction solutionwas adjusted to 8 using triethylamine, the solvent was distilled offunder reduced pressure and the residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/20, 1/3) toobtain 59 (1.91 g, 91%) as a pink oily substance.

APCI-MS m/z 252[M+H]⁺

Synthesis of 60

To a 1,2-dimethoxyethane (62 ml) solution of 59 (1.90 g, 7.55 mmol) and6 (2.08 g, 7.55 mmol), potassium carbonate (3.13 g, 22.65 mmol), water(1.31 ml) and tetrakistriphenylphosphine palladium (436 mg, 0.38 mmol)were added under an argon atmosphere, and the mixture was stirred at 80°C. for 27 hours. The reaction solution was allowed to return to roomtemperature and water was added, and the solution was extracted withethyl acetate. The extraction liquid was washed with water and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/19) and then recrystallized from ethylacetate-n-hexane (1/9) to obtain 60 (1.50 g, 54%) as yellow crystals.

mp 86-87° C.

APCI-MS m/z 365[M+H]⁺

Synthesis of 61

To a methylene chloride (12 ml) solution of 60 (1.50 g, 4.12 mmol),trifluoroacetic acid (8 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 15minutes. To the reaction solution, ice water and ethyl acetate wereadded and extracted with ethyl acetate after adjusting the pH to 9 usingan aqueous potassium carbonate solution. The extraction liquid waswashed with water and dried, and then the solvent was distilled offunder reduced pressure. The residue was recrystallized from ethylacetate-n-hexane to obtain 61 (1.13 g, 94%) as yellow crystals.

mp 223-224° C.

APCI-MS m/z 293[M+H]⁺

Synthesis of THK-5072

To a mixture of 61 (500 mg, 1.71 mmol), 62 (690 mg, 2.05 mmol),triphenylphosphine (540 mg, 2.05 mmol) and tetrahydrofuran (25 ml),diisopropyl azodicarboxylate (0.41 ml, 2.05 mmol) was added dropwiseunder ice cooling and stirring, and the mixture was stirred at roomtemperature for 16 hours. To the reaction solution, 62 (690 mg, 2.05mmol), triphenylphosphine (540 mg, 2.05 mmol) and tetrahydrofuran (5 ml)were added under ice cooling and stirring, and diisopropylazodicarboxylate (0.41 ml, 2.05 mmol) was added dropwise, followed byfurther stirring at room temperature for 6 hours. The solvent of thereaction solution was distilled off under reduced pressure and theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/4) to obtain a pale yellow oilysubstance (1.47 g). To an anisole (4 ml) solution of the presentproduct, methanesulfonic acid (12 ml) was added dropwise under icecooling and stirring, followed by stirring at room temperature for 15minutes. The reaction solution was poured into ice water and ethylacetate was added, and the solution was extracted with ethyl acetateafter adjusting the pH to 9 using an aqueous potassium carbonatesolution. The extraction liquid was washed with water and dried, andthen the solvent was distilled off under reduced pressure. The residuewas purified by silica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/9, 1/1) to obtain THK-5072 (620 mg, 69%) as a yellowfoam-like substance.

¹H NMR (400 MHz, DMSO-d₆) δ 1.15 (6H, t, J=7.3 Hz), 2.33 (3H, s), 3.43(4H, q, J=7.3 Hz), 3.57-3.71 (2H, m), 4.30 (1H, dd, J=11, 5.7 Hz), 4.38(1H, dd, J=11, 3.0 Hz), 4.71-4.77 (1H, m), 5.11 (1H, t, J=5.6 Hz), 6.79(2H, d, J=9.1 Hz), 7.01 (1H, dd, J=8.8, 2.4 Hz), 7.32 (1H, d, J=2.1 Hz),7.39 (2H, d, J=8.2 Hz), 7.75 (2H, d, J=8.5 Hz), 7.78 (1H, d, J=8.8 Hz),7.85 (1H, d, J=8.5 Hz), 8.10 (2H, d, J=9.1 Hz), 8.21 (1H, d, J=8.8 Hz)

IR (Nujol) 1619, 1593 cm⁻¹,

APCI-MS m/z 521[M+H]⁺

Synthesis Method of THK-5073

Synthesis of 64

To a tetrahydrofuran (45 ml) solution of 61 (630 mg, 2.16 mmol), 63 (932mg, 2.59 mmol) and triphenylphosphine (678 mg, 2.59 mmol), diisopropylazodicarboxylate (0.513 ml, 2.59 mmol) was added dropwise under icecooling and stirring, and the mixture was stirred at room temperaturefor 16 hours. To the reaction solution, 63 (788 mg, 2.18 mmol),triphenylphosphine (573 mg, 2.18 mmol) and tetrahydrofuran (5 ml) wereadded under ice cooling and stirring, and diisopropyl azodicarboxylate(0.433 ml, 2.18 mmol) was added dropwise, followed by further stirringat room temperature for 3 days. The solvent of the reaction solution wasdistilled off under reduced pressure and the residue was purified bysilica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/9) to obtain 64 (720 mg, 53%) as a pale yellowfoam-like substance.

APCI-MS m/z 635[M+H]⁺

Synthesis of 65

To a methylene chloride (12 ml) solution of 64 (710 mg, 1.12 mmol),trifluoroacetic acid (8 ml) was added dropwise under ice cooling andstirring, and water (2 ml) was added and the mixture was stirred at roomtemperature for 5.5 hours. To the reaction solution, ice water and ethylacetate were added and the solution was extracted with ethyl acetateafter adjusting the pH to 8 using an aqueous saturated sodium hydrogencarbonate solution. The extraction liquid was washed with water anddried, and then the solvent was distilled off under reduced pressure.The residue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/4, 1/2) to obtain 65 (520 mg, 85%) asa yellow foam-like substance.

APCI-MS m/z 221[M+H]⁺

Synthesis of THK-5073

To a methylene chloride (60 ml) solution of 65 (290 mg, 0.557 mmol),3,4-dihydro-2H-pyran (0.51 ml, 5.6 mmol) and a paratoluenesulfonic acidpyridine salt (182 mg, 0.724 mmol) were added, and the mixture wasstirred at room temperature for 3 days. To the reaction solution,paratoluenesulfonic acid monohydrate (125 mg, 0.724 mmol) and3,4-dihydro-2H-pyran (0.51 ml, 5.6 mmol) were added, followed by furtherstirring at room temperature for 1 hour. The reaction solution wasneutralized with triethylamine and the solvent was distilled off underreduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/3, 1/2) toobtain THK-5073 (336 mg, 100%) as a yellow oily substance. ¹H NMR (500MHz, DMSO-d₆) δ 1.15(6H, t, J=7.2 Hz), 1.36-1.74 (7H, m), 2.34 (3H, s),3.43 (4H, q, J=7.2 Hz), 3.69-3.75, 3.84-3.90 (1H, m), 4.13-4.37 (5H, m),4.71-4.76, 4.86-4.88 (1H, m), 6.79 (2H, d, J=9.0 Hz), 7.04 (1H, dd,J=9.0, 1.9 Hz), 7.25-7.29 (1H, m), 7.39-7.44 (2H, m), 7.77-7.82 (3H, m),7.86 (1H, d, J=8.7 Hz), 8.11 (2H, d, J=8.7 Hz), 8.19-8.25 (1H, m)

APCI-MS m/z 605[M+H]⁺

Synthesis Method of THK-5077

Synthesis of 67

To a mixture of 34 (1.50 g, 3.3 mmol), 66 (1.2 g, 3.6 mmol) and1,2-dimethoxyethane (15 ml), an aqueous 2M sodium carbonate solution(3.3 ml, 6.6 mmol) and tetrakistriphenylphosphine palladium (189 mg,0.16 mmol) were added under an argon atmosphere, and the mixture wasstirred at 90° C. for 1.5 hours. The reaction solution was allowed toreturn to room temperature and insolubles were removed by filtration,and the solution was washed with chloroform. The filtrate and the washwere combined and the mixture was washed with an aqueous saturatedsodium hydrogen carbonate solution and dried, and then the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel flash column chromatography (eluting solvent: ethylacetate/n-hexane=1/9, 1/4, 1/2) to obtain 67 (1.70 g, 100%) as acolorless oily substance.

APCI-MS m/z 517[M+H]⁺

Synthesis of 68

To a methylene chloride (8 ml) solution of 67 (1.70 g, 3.29 mmol),trifluoroacetic acid (2 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 16 hours.The solvent of the reaction solution was distilled off under reducedpressure and chloroform was added. The solution was made basic with anaqueous saturated sodium hydrogen carbonate solution and potassiumcarbonate and extracted with chloroform. The extraction liquid was driedand the solvent was distilled off under reduced pressure, and then theresidue was purified by NH silica gel flash column chromatography(eluting solvent: n-hexane/ethyl acetate=3/1) to obtain 68 (1.34 g, 97%)as a yellow oily substance.

APCI-MS m/z 417[M+H]⁺

Synthesis of THK-5077

To a mixture of 68 (101 mg, 0.243 mmol) and thioanisole (2 ml),methanesulfonic acid (0.4 ml) was added under ice cooling and stirring,and the mixture was stirred at room temperature for 2 hours. To thereaction solution, ice water was added and the solution washed withethyl acetate. The ethyl acetate layer was extracted with water, and theextract layer and the previous aqueous layer were combined. The mixturewas made basic with potassium carbonate and extracted with chloroform.The extraction liquid was dried and the solvent was distilled off underreduced pressure, and then the residue was purified by NH silica gelflash column chromatography (eluting solvent: n-hexane/ethylacetate=2/1, 1/1, 1/2) and then recrystallized frommethanol-diisopropylether to obtain THK-5077 (54 mg, 68%) as a yellowsolid.

mp 143.5-145.5° C., ¹H NMR (400 MHz, DMSO-d₆) δ 2.75(3H, d, J=4.9 Hz),3.6-3.8 (2H, m), 4.6 to 4.9 (3H, m), 5.10 to 5.13 (1H, m), 5.12 (1H, q,J=4.9 Hz), 6.10 (1H, brs), 6.66 (2H, d, J=9.0 Hz), 7.16 (1H, dd, J=8.8,2.4 Hz), 7.45 (1H, d, J=2.4 Hz), 7.81 (1H, d, J=8.9 Hz), 7.84 (1H, d,J=8.8 Hz), 8.07 (2H, d, J=9.0 Hz), 8.20 (1H, d, J=7.5 Hz)

IR (Nujol) 2854, 1612, 1461 cm⁻¹

APCI-MS m/z 327[M+H]⁺

Synthesis Method of THK-5078

Synthesis of 69

A mixture of 68 (637 mg, 1.50 mmol), acetoaldehyde (115 mg, 2.60 mmol),a picoline borane complex (279 mg, 2.60 mmol) and methanol (10ml)-acetic acid (1 ml) was stirred at room temperature for 16 hours.Acetoaldehyde (1.15 g, 26 mmol) and a picoline borane complex (279 mg,2.60 mmol) were added, followed by further stirring at room temperaturefor 16 hours. The solvent of the reaction solution was distilled offunder reduced pressure and aqueous 10% hydrochloric acid (10 ml) wasadded to the residue. After stirring at room temperature for 30 minutes,the solution was made basic by adding potassium carbonate. The reactionsolution was extracted with chloroform and the extraction liquid wasdried, and the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel flash column chromatography (elutingsolvent: n-hexane/ethyl acetate=4/1) to obtain 69 (640 mg, 94%) as ayellow oily substance.

APCI-MS m/z 445[M+H]⁺

Synthesis of THK-5078

To a mixture of 69 (639 mg, 1.44 mmol) and thioanisole (5 ml),methanesulfonic acid (1 ml) was added under ice cooling and stirring,and the mixture was stirred at room temperature for 1 hour. To thereaction solution, ice water was added and the solution was washed withethyl acetate. The ethyl acetate layer was extracted with water, and theextraction layer and the previous aqueous layer were combined. Themixture was made basic with potassium carbonate and extracted withchloroform. The extraction liquid was dried and the solvent wasdistilled off under reduced pressure, and then the residue was purifiedby NH silica gel flash column chromatography (eluting solvent:n-hexane/ethyl acetate=2/1, 1/1) and then an oxalate was formed inacetone to obtain THK-5078 (460 mg, 71%) as orange crystals.

mp 134-136° C., ¹H NMR (400 MHz, DMSO-d₆) δ 1.10 (3H, t, J=7.0 Hz), 3.49(2H, q, J=7.0 Hz), 3.7-3.8 (2H, m), 4.6-4.9 (3H, m), 6.83 (2H, d, J=9.0Hz), 7.20 (1H, dd, J=8.8, 2.4 Hz), 7.49 (1H, d, J=2.4 Hz), 7.85 (1H, d,J=9.0 Hz), 7.89 (1H, d, J=7.3 Hz), 8.11 (2H, d, J=9.0 Hz), 8.26 (1H, d,J=9.0 Hz)

IR (Nujol) 1603, 1458, 1214 cm⁻¹

APCI-MS m/z 355[M+H]⁺

Synthesis Method of THK-5090

Synthesis of 70

To a tetrahydrofuran (60 ml) solution of 58 (1.36 g, 7.57 mmol), 22(2.48 g, 9.09 mmol) and triphenylphosphine (2.38 g, 9.09 mmol),diisopropyl azodicarboxylate (1.8 ml, 9.09 mmol) was added dropwiseunder ice cooling and stirring, and the mixture was stirred at roomtemperature for 16 hours. To the reaction solution, 22 (1.24 g, 4.55mmol), triphenylphosphine (1.19 g, 4.55 mmol) and tetrahydrofuran (10ml) were added under ice cooling and stirring, and diisopropylazodicarboxylate (0.9 ml, 4.55 mmol) was added dropwise, followed byfurther stirring at room temperature for 4 hours. The solvent of thereaction solution was distilled off under reduced pressure and theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/9) to obtain 70 (3.04 g, 93%) as apale pink oily substance.

APCI-MS m/z 434[M+H]⁺

Synthesis of 71

To a 1,2-dimethoxyethane (40 ml) solution of 70 (2.00 g, 4.61 mmol) and56 (1.14 g, 4.61 mmol), potassium carbonate (1.91 g, 13.83 mmol), water(0.8 ml) and tetrakistriphenylphosphine palladium (270 mg, 0.23 mmol)were added under an argon atmosphere, and the mixture was stirred at 80°C. for 2 days. The reaction solution was allowed to return to roomtemperature and water was added, and the solution was extracted withethyl acetate. The extraction liquid was washed with water and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/9) to obtain 71 (1.22 g, 51%) as ayellow oily substance.

APCI-MS m/z 519 [M+H]+

Synthesis of 72

To a mixture of 71 (1.21 g, 2.33 mmol) and anisole (3 ml),methanesulfonic acid (9 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 30minutes. The reaction solution was poured into ice water and thesolution was extracted with ethyl acetate-tetrahydrofuran. Theextraction liquid was dried and the solvent was distilled off underreduced pressure, and then the residue was washed with ethylacetate-n-hexane (1/2) to obtain 72 (680 mg, 86%) as yellow crystals.

mp 193-194° C.

APCI-MS m/z 339[M+H]⁺

Synthesis of THK-5090

To a pyridine (15 ml) solution of 72 (690 mg, 2.04 mmol), atetrahydrofuran (15 ml) solution of paratoluenesulfonic anhydride (1.00g, 3.06 mmol) was added dropwise over 35 minutes under ice cooling andstirring, and the mixture was stirred at the same temperature for 30minutes. To the reaction solution, ice water was added and the solutionwas extracted with ethyl acetate after adjusting the pH to 9 using anaqueous potassium carbonate solution. The extraction liquid was driedand the solvent was distilled off under reduced pressure (tolueneazeotrope). The residue was purified by silica gel column chromatography(eluting solvent: ethyl acetate/n-hexane=1/2, 1/1) and thenrecrystallized from ethyl acetate-n-hexane to obtain THK-5090 (459 mg,46%) as pale yellow crystals.

mp 100-102° C., ¹H NMR (400 MHz, DMSO-d₆) δ 2.33 (3H, s), 3.03 (6H, s),3.56-3.71 (2H, m), 4.30 (1H, dd, J=11, 6.0 Hz), 4.39 (1H, dd, J=11, 2.7Hz), 4.72-4.78 (1H, m), 5.12 (1H, t, J=5.4 Hz), 6.84 (2H, d, J=9.1 Hz),7.02 (1H, dd, J=9.1, 2.4 Hz), 7.33 (1H, d, J=2.4 Hz), 7.39 (2H, d, J=7.9Hz), 7.75 (2H, d, J=8.2 Hz), 7.79 (1H, d, J=9.1 Hz), 7.88 (1H, d, J=8.5Hz), 8.13 (2H, d, J=9.1 Hz), 8.23 (1H, d, J=7.9 Hz)

IR (Nujol) 1731, 1609, 1597 cm⁻¹

APCI-MS m/z 493[M+H]⁺

Synthesis Method of THK-5075

Synthesis of THK-5075

A mixture of 73 (1.19 g, 8.74 mmol), 74 (1.06 ml, 8.74 mmol), potassiumhydroxide (590 mg, 10.5 mmol) and toluene (40 ml) was heated at refluxfor 16 hours. The reaction solution was allowed to return to roomtemperature, and water and an aqueous saturated ammonium chloridesolution were added and the solution was extracted with ethyl acetate.Insolubles were removed by filtration with celite. The extraction liquidwas washed with water and dried, and then the solvent was distilled offunder reduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: n-hexane/ethyl acetate=9/1, 2/1) andthen recrystallized from ethyl acetate-n-hexane to obtain THK-5075 (737mg, 35%) as pale yellow crystals.

mp 160-161° C., ¹H NMR (500 MHz, DMSO-d₆) δ 5.80 (2H, s), 6.96-7.00 (2H,m), 7.33 (2H, t, J=8.8 Hz), 7.63 (1H, d, J=9.6 Hz), 7.68 (1H, d, J=8.4Hz), 8.11 (1H, d, J=8.4 Hz), 8.24 (2H, dd, J=9.0, 5.5 Hz)

IR (Nujol) 3388, 1619, 1600 cm⁻¹

APCI-MS m/z 239[M+H]⁺

Synthesis Method of THK-5076

Synthesis of 76

To an ethanol (20 ml) suspension of 75 (1.00 g, 5.15 mmol), an ironpowder (1.15 g) and 0.1N hydrochloric acid (2.58 ml) were added and themixture was heated at reflux for 16 hours, and then 0.1N hydrochloricacid (7.72 ml) were added, followed by further heating at reflux for 6hours. To the reaction solution, 74 (0.625 ml, 5.15 mmol) and potassiumhydroxide (0.347 g, 6.18 mmol) were added, and the mixture was heated atreflux for 3 days. The reaction solution was allowed to return to roomtemperature and insolubles were removed by filtration, and the solutionwas washed with methanol. The filtrate and the wash were combined andthe solvent was distilled off under reduced pressure. Water was added tothe residue and the solution was extracted with ethyl acetate. The ethylacetate layer was extracted with aqueous 10% hydrochloric acid, and thehydrochloric acid extraction liquid was extracted with ethyl acetateafter adjusting the pH to 9 using an aqueous potassium carbonatesolution. The extraction liquid was washed with water and dried, andthen the solvent was distilled off under reduced pressure. The residuewas purified by silica gel column chromatography (eluting solvent:n-hexane/ethyl acetate=9/1, 4/1) to obtain 76 (225 mg, 16%) as a yellowoily substance.

APCI-MS m/z 267[M+H]⁺

Synthesis of THK-5076

To a methanol (20 ml) solution of 76 (540 mg, 2.03 mmol), 4Mhydrochloric acid/ethyl acetate (1.02 ml) was added dropwise and thesolvent was distilled off under reduced pressure. The residue wasrecrystallized from ethyl acetate-isopropanol to obtain THK-5076 (478mg) as orange crystals.

mp 250-251° C., ¹H NMR (400 MHz, DMSO-d₆) δ 3.26 (6H, s), 7.46-7.57 (4H,m), 7.85 (1H, d, J=8.2 Hz), 8.06 (1H, d, J=9.4 Hz), 8.23 to 8.29 (2H,m), 8.77 (1H, d, J=8.2 Hz), 15.0 (1H, br)

IR (Nujol) 1634, 1599 cm⁻¹

APCI-MS m/z 267[M+H]⁺

Synthesis Method of THK-5079

Synthesis of 78

To a chloroform (65 ml) solution of 77 (5.00 g, 28.71 mmol), achloroform (27 ml)-methanol (7 ml) solution of metachlorobenzoic acid(10.9 g, 63.16 mmol) was added dropwise at room temperature, and themixture was stirred at room temperature for 16 hours. To the reactionsolution, an aqueous sodium thiosulfate solution was added, and themixed solution was stirred room temperature for 10 minutes and extractedwith chloroform. The extraction liquid was washed with an aqueouspotassium carbonate solution, purified as it is by silica gel columnchromatography (eluting solvent: chloroform) and then washed withn-hexane-ethyl acetate (2/1) to obtain 78 (4.96 g, 91%) as yellowcrystals.

mp 164-165° C.

APCI-MS m/z 191[M+H]⁺

Synthesis of 79

To 78 (4.94 g, 26 mmol), phosphorus oxyschloride (22 ml) was addeddropwise under ice cooling and stirring, and the mixture was heated atreflux for 3 hours. The reaction solution was ice-cooled and ice waterwas added, and then the solution was made basic with ammonia water andextracted with chloroform. The extraction liquid was washed withsaturated saline and dried, and then the solvent was distilled off underreduced pressure. The residue was purified by silica gel flash columnchromatography (eluting solvent: n-hexane/methylene chloride=2/1, 1/1,1/2) and then washed with n-hexane to obtain 79 (2.25 g, 42%) as a paleyellow solid.

APCI-MS m/z 209/211[M+H]⁺

Synthesis of 81

To a mixture of 79 (1.04 g, 5 mmol), 80 (770 mg, 5.5 mmol) and1,2-dimethoxyethane (25 ml), an aqueous 2M sodium carbonate solution (5ml, 10 mmol) and tetrakistriphenylphosphine palladium (289 mg, 0.25mmol) were added under an argon atmosphere, and the mixture was stirredat 90° C. for 3 hours. The reaction solution was allowed to return toroom temperature and the solvent was distilled off under reducedpressure. To the residue, chloroform was added and insolubles wereremoved by filtration, and the solution was washed with chloroform. Thefiltrate and the wash were combined and the mixture was washed withsaturated saline and dried, and then the solvent was distilled off underreduced pressure. The residue was purified by silica gel flash columnchromatography (eluting solvent: methylene chloride/n-hexane=1/1) andthen washed with n-hexane to obtain 81 (1.30 g, 97%) as a pale yellowsolid.

APCI-MS m/z 269 [M+H]+

Synthesis of THK-5079

A mixture of 81 (1.29 g, 4.81 mmol), 10% Pd—C (130 mg) and ethanol (30ml) was stirred under a hydrogen atmosphere at room temperature for 2hours. The catalyst was removed by filtration and the solvent of thefiltrate was distilled off under reduced pressure, and then the residuewas washed with n-hexane to obtain THK-5079 (1.087 g, 95%) as ayellowish orange solid.

mp 105-108° C., ¹H NMR (400 MHz, DMSO-d₆) δ 5.9-6.3 (2H, brs), 6.70 (1H,dd, J=8, 1 Hz), 7.23 (1H, d, J=8 Hz), 7.36 (2H, t, J=9 Hz), 7.44 (1H, t,J=8 Hz), 7.98 (1H, d, J=9 Hz), 8.31 (2H, dd, J=9, 5 Hz), 8.63 (1H, d,J=9 Hz)

IR (Nujol) 1614, 1592 cm⁻¹

APCI-MS m/z 239[M+H]⁺

Synthesis Method of THK-5080

Synthesis of 82

A mixture of THK-5079 (450 mg, 1.9 mmol), an aqueous 35% formalinsolution (0.81 g, 9.4 mmol), a picoline borane complex (303 mg, 2.8mmol) and methanol (10 ml)-acetic acid (1 ml) was stirred at roomtemperature for 30 minutes and a picoline borane complex (100 mg, 0.93mmol) was added, followed by stirring at room temperature for 30minutes. The solvent of the reaction solution was distilled off underreduced pressure and aqueous 10% hydrochloric acid (5 ml) was added tothe residue. After stirring at room temperature for 30 minutes, thesolution was made basic by adding potassium carbonate. The reactionsolution was extracted with chloroform, and the extraction liquid wasdried and the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel flash column chromatography (elutingsolvent:

n-hexane/ethyl acetate=49/1, 19/1) to obtain 82 (465 mg, 92%) as agreenish yellow oily substance.

APCI-MS m/z 267[M+H]⁺

Synthesis of THK-5080

After dissolving by adding oxalic acid (312 mg, 3.47 mmol) to an acetone(10 ml) solution of 82 (462 mg, 1.73 mmol), the solvent was distilledoff under reduced pressure. The residue was washed with diisopropyletherto obtain THK-5080 (620 mg, 89%) as an orange solid.

mp 118-120° C., ¹H NMR (500 MHz, DMSO-d₆) δ 2.87 (2H, s), 7.15 (1H, dd,J=6, 1.5 Hz). 7.39 (2H, t, J=9 Hz), 7.6-7.7 (2H, m), 8.11 (1H, d, J=9Hz), 8.33 (2H, dd, J=9, 5 Hz), 8.59 (1H, d, J=9 Hz)

IR (Nujol) 1638, 1603 cm⁻¹

APCI-MS m/z 267[M+H]⁺

Synthesis Method of THK-5081

Synthesis of 84

A mixture of 83 (4.78 g, 29.84 mmol) and Boc₂O (10.28 ml, 44.8 mmol) wasstirred at 80° C. for 2 hours, at 100° C. for 1 hour, followed bystirring at 120° C. for 30 minutes. The reaction solution was purifiedby silica gel column chromatography (eluting solvent: n-hexane/ethylacetate=4/1, 1/1) to obtain 84 (6.54 g, 84%) as a yellow solid.

mp 126-128° C.

APCI-MS m/z 261[M+H]⁺

Synthesis of 85

To a chloroform (120 ml) solution of 84 (6.62 g, 25.43 mmol),paratoluenesulfonyl chloride (5.82 g, 30.5 mmol) and potassium carbonate(4.22 g, 30.5 mmol) were added at room temperature under stirring, andthe mixture was heated at reflux for 5 hours. The reaction solution wasallowed to return to room temperature and water was added, and thesolution was extracted with chloroform. The extraction liquid waspurified as it is by NH silica gel column chromatography (elutingsolvent: chloroform) and then silica gel column chromatography (elutingsolvent: n-hexane/ethyl acetate=100/1 to ethyl acetate) to obtain 85(3.70 g, 52%) as a pale yellow solid.

mp 99-100° C.

APCI-MS m/z 279[M+H]⁺

Synthesis of 87

To a 1,2-dimethoxyethane (44 ml) solution of 85 (1.50 g, 5.38 mmol) and86 (1.20 g, 5.38 mmol), potassium carbonate (2.23 g, 16.13 mmol), water(0.94 ml) and tetrakistriphenylphosphine palladium (310 mg, 0.27 mmol)were added under an argon atmosphere, and the mixture was stirred at 80°C. for 3 hours. The reaction solution was allowed to return to roomtemperature and water was added, and the solution was extracted withethyl acetate. The extraction liquid was washed with water and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/500, 1/100) to obtain 87 (1.68 g, 92%)as a colorless solid.

APCI-MS m/z 339 [M+H]

Synthesis of THK-5081

To a chloroform (12 ml) solution of 87 (1.50 g, 4.43 mmol),trifluoroacetic acid (8 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 3 hours.To the reaction solution, water was added and the solution was extractedwith ethyl acetate after adjusting the pH to 9 using an aqueouspotassium carbonate solution. The extraction liquid was washed withwater and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: ethyl acetate/n-hexane=1/9) and then recrystallizedfrom n-hexane to obtain THK-5081 (404 mg, 38%) as pale yellow crystals.

mp 87.5-88° C., ¹H NMR (500 MHz, DMSO-d₆) δ 6.25 (2H, br), 6.91 (1H, d,J=7.7 Hz), 7.09 (1H, d, J=8.0 Hz), 7.29 (1H, t, J=8.0 Hz), 7.35 (2H, t,J=9.0 Hz), 8.08 (1H, d, J=8.7 Hz), 8.26 (1H, d, J=8.7 Hz), 8.43 (2H, dd,J=8.7, 5.5 Hz)

IR (Nujol) 3433, 1616, 1600 cm⁻¹

APCI-MS m/z 239[M+H]⁺

Synthesis Method of THK-5082

Synthesis of THK-5082

To a methanol (21 ml)-acetic acid (2.1 ml) solution of THK-5081 (450 mg,1.9 mmol) and an aqueous 35% formalin solution (1.51 ml, 18.9 mmol), apicoline borane complex (605 mg, 5.67 mmol) was added under ice coolingand stirring, and the mixture was stirred at room temperature for 16hours. To the reaction solution, an aqueous potassium carbonate solutionwas added thereby adjusting the pH to 8, and the solution was extractedwith ethyl acetate. The extraction liquid was washed with water anddried, and then the solvent was distilled off under reduced pressure.The residue was purified by silica gel column chromatography (elutingsolvent: n-hexane/ethyl acetate=50/1) and then NH silica gel columnchromatography (eluting solvent: n-hexane/ethyl acetate=100/1) to obtaina free form (430 mg) of THK-5082 as a yellow oily substance. The presentproduct was dissolved in ethyl acetate (10 ml) and 4M hydrochloricacid/ethyl acetate (0.4 ml) was added dropwise, followed by stirring atroom temperature for 30 minutes. The precipitate was collected byfiltration and dried to obtain THK-5082 (430 mg, 75%) as colorlesscrystals.

mp 216-216.5° C., ¹H NMR (400 MHz, DMSO-d₆) δ 3.57 (6H, s), 7.41-7.48(2H, m), 7.76 (1H, t, J=7.9 Hz), 8.15 (1H, d, J=7.9 Hz), 8.24 (1H, d,J=7.6 Hz), 8.38 (1H, d, J=8.8 Hz), 8.56-8.62 (2H, m), 8.66 (1H, d, J=8.8Hz), 12.2 (1H, br)

IR (Nujol) 2252, 1602 cm⁻¹

APCI-MS m/z 267 [M+H]⁺

Synthesis Method of THK-5086

Synthesis of 89

To a methanol (50 ml)-tetrahydrofuran (50 ml) suspension of 88 (5.00 g,28.71 mmol), 10% Pd—C (moisture of about 50%; 1.00 g) was added under ahydrogen atmosphere, and the mixture was stirred at room temperature for2 hours. The catalyst was removed by filtration and the solvent of thefiltrate was distilled off under reduced pressure to obtain 89 (4.14 g,100%) as a pale yellow solid.

mp 116-117° C.

APCI-MS m/z 145[M+H]⁺

Synthesis of 90

A mixture of 89 (4.14 g, 28.71 mmol) and Boc₂O (9.9 ml, 43.1 mmol) wasstirred at 120° C. for 30 minutes. To the reaction solution, silica gel(30 ml) and toluene (100 ml) were added, followed by stirring at 80° C.for 1 hour. The reaction solution was allowed to return to roomtemperature and purified as it is by silica gel column chromatography(eluting solvent: n-hexane/ethyl acetate=2/1, ethyl acetate) to obtain90 (5.92 g, 84%) as a pale yellow solid.

mp 132-133° C.

APCI-MS m/z 245 [M+H]⁺

Synthesis of 91

To a chloroform (55 ml) solution of 90 (5.91 g, 24.19 mmol), achloroform (22 ml)-methanol (6 ml) solution of metachlorobenzoic acid(9.19 g, 53.2 mmol) was added dropwise at room temperature, and themixed solution was stirred at room temperature for 16 hours. To thereaction solution, an aqueous sodium thiosulfate solution was added,followed by stirring at room temperature for 10 minutes and furtherextraction with chloroform. The extraction liquid was washed with anaqueous potassium carbonate solution, purified as it is by silica gelcolumn chromatography (eluting solvent: chloroform) and then washed withn-hexane-ethyl acetate (2/1) to obtain 91 (5.25 g, 83%) as colorlesscrystals.

mp 212-213° C.

APCI-MS m/z 261 [M+H]⁺

Synthesis of 92

To a chloroform (200 ml) solution of 91 (5.25 g, 20.17 mmol),paratoluenesulfonyl chloride (4.61 g, 24.2 mmol) and potassium carbonate(3.35 g, 24.2 mmol) were added at room temperature under stirring, andthe mixture was heated at reflux for 3.5 hours. The reaction solutionwas allowed to return to room temperature and water was added, and thesolution was extracted with chloroform. The extraction liquid waspurified as it is by NH silica gel column chromatography (elutingsolvent: chloroform) and then silica gel column chromatography (elutingsolvent: n-hexane/ethyl acetate=9/1, 4/1) to obtain 92 (1.87 g, 33%) asa colorless solid.

mp 152-153° C.

APCI-MS m/z 279/281[M+H]⁺

Synthesis of 93

To a 1,2-dimethoxyethane (30 ml) solution of 92 (1.00 g, 3.59 mmol) and80 (0.60 g, 4.31 mmol), potassium carbonate (1.50 g, 10.8 mmol), water(0.62 ml) and tetrakistriphenylphosphine palladium (210 mg, 0.18 mmol)were added under an argon atmosphere, and the mixture was stirred at 80°C. for 5.5 hours. The reaction solution was allowed to return to roomtemperature and water was added, and the solution was extracted withethyl acetate. The extraction liquid was dried and the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/4)and then recrystallized from ethyl acetate-n-hexane to obtain 93 (990mg, 82%) as colorless crystals.

mp 194-195° C.

APCI-MS m/z 339[M+H]⁺

Synthesis of THK-5086

To a chloroform (12 ml) suspension of 93 (980 mg, 2.90 mmol),trifluoroacetic acid (8 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 1.5 hours.To the reaction solution, water was added and the solution was extractedwith ethyl acetate after adjusting the pH to 9 using an aqueouspotassium carbonate solution. The extraction liquid was washed withwater and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: ethyl acetate/n-hexane=1/2) and then recrystallizedfrom ethyl acetate-n-hexane to obtain THK-5086 (635 mg, 92%) as paleyellow crystals.

mp 145.5-146° C., ¹H NMR (500 MHz, DMSO-d₆) δ 5.71 (2H, s), 6.82 (1H, d,J=2.6 Hz), 7.19 (1H, dd, J=9.0, 2.6 Hz), 7.29-7.35 (2H, m), 7.75 (1H, d,J=9.0 Hz), 7.89 (1H, d, J=8.7 Hz), 8.03 (1H, d, J=8.7 Hz), 8.19-8.24(2H, m)

IR (Nujol) 3342, 1628 cm⁻¹

APCI-MS m/z 239[M+H]⁺

Synthesis Method of THK-5087

Synthesis of THK-5087

To a methanol (21 ml)-acetic acid (2.1 ml) solution of THK-5086 (440 mg,1.85 mmol) and an aqueous 37% formalin solution (1.48 ml, 18.5 mmol), apicoline borane complex (590 mg, 5.55 mmol) was added under ice coolingand stirring, and the mixture was stirred at room temperature for 3.5hours. After adjusting the pH of the reaction solution to 9 by adding anaqueous potassium carbonate solution, the solution was extracted withethyl acetate. The extraction liquid was washed with water and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: n-hexane/ethyl acetate=9/1, 4/1) and then recrystallized fromn-hexane-ethyl acetate to obtain THK-5087 (415 mg, 84%) as pale yellowcrystals.

mp 172-173° C., ¹H NMR (400 MHz, DMSO-d₆) δ 3.06(6H, s), 6.96 (1H, d,J=2.7 Hz), 7.30-7.37 (2H, m), 7.47 (1H, dd, J=9.4, 3.0 Hz), 7.88 (1H, d,J=9.4 Hz), 7.96 (1H, d, J=8.8 Hz), 8.17 (1H, d, J=8.8 Hz), 8.22-8.28(2H, m)

IR (Nujol) 1618 cm⁻¹

APCI-MS m/z 267[M+H]⁺

Synthesis Method of THK-932

Synthesis of 95

To a 1,2-dimethoxyethane (27 ml) solution of 34 (1.05 g, 2.28 mmol) and94 (720 mg, 2.28 mmol), potassium carbonate (940 mg, 6.83 mmol), water(0.57 ml) and tetrakistriphenylphosphine palladium (131 mg, 0.114 mmol)were added under an argon atmosphere, and the mixture was stirred at 80°C. for 1.5 hours. The reaction solution was allowed to return to roomtemperature, diluted with ethyl acetate, washed with water and thendried. The solvent was distilled off under reduced pressure and theresidue was purified by silica gel column chromatography (elutingsolvent: n-hexane/ethyl acetate=4/1→3/1) to obtain 95 (900 mg, 78%) as apale yellow solid.

mp 150-152° C., APCI-MS m/z 504[M+H]⁺

Synthesis of 96

To a chloroform (12 ml) solution of 95 (900 mg, 1.79 mmol),trifluoroacetic acid (8 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 2 hours.The reaction solution was ice-cooled, neutralized with an aqueouspotassium carbonate solution and extracted with ethyl acetate. Theorganic layer was washed with water and dried, and then solvent wasconcentrated under reduced pressure to obtain 96 (720 mg, 99%) as a paleyellow oily substance.

APCI-MS m/z 404[M+H]⁺

Synthesis of THK-932

To an anisole (5 ml) solution of 96 (720 mg, 1.78 mmol), methanesulfonicacid (20 ml) was added dropwise under ice cooling and stirring, and themixture was stirred at room temperature for 30 minutes. The reactionsolution was ice-cooled and ice water was added dropwise, and thesolution was washed with ethyl acetate. The aqueous layer was extractedwith ethyl acetate after adjusting the pH to 9 using an aqueouspotassium carbonate solution. The organic layer was washed with waterand dried, and the solvent was distilled off under reduced pressure. Theresidue was purified by NH silica gel column chromatography (elutingsolvent: n-hexane/ethyl acetate=1/1→ethyl acetate) and thenrecrystallized from ethyl acetate to obtain THK-932 (390 mg, 69%) ascolorless crystals.

mp 160-161° C., ¹H NMR (500 MHz, DMSO-d₆) δ 3.66-3.77 (2H, m), 4.65-4.86(3H, m), 5.14 (1H, t, J=5.8 Hz), 6.41 (2H, s), 6.57 (1H, d, J=9.0 Hz),7.20 (1H, dd, J=8.8, 2.4 Hz), 7.48 (1H, d, J=2.4 Hz), 7.84 (1H, d, J=8.8Hz), 7.87 (1H, d, J=8.8 Hz), 8.18-8.38 (2H, m), 8.83 (1H, d, J=2.4 Hz)

IR (Nujol) 3335, 1638, 1613 cm⁻¹

APCI-MS m/z 314[M+H]⁺

Synthesis Method of THK-5088

Synthesis of 98

To a mixture of 97 (387 mg, 2.17 mmol), acetoaldehyde (2.39 g, 54.3mmol) and methanol (10 ml)-acetic acid (1 ml), a picoline borane complex(1.16 g, 10.8 mmol) was added under ice cooling and stirring, and themixture was stirred at room temperature for 3.5 hours. The solvent ofthe reaction solution was distilled off under reduced pressure andaqueous 10% hydrochloric acid (5 ml) was added to the residue. Afterstirring at room temperature for 30 minutes, the solution was made basicby adding potassium carbonate. The reaction solution was extracted withchloroform and the extraction liquid was dried, and the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel flash column chromatography (eluting solvent: n-hexane/ethylacetate=19/1) to obtain 98 (353 mg, 69%) as a yellow oily substance.

APCI-MS m/z 235/237[M+H]⁺

Synthesis of THK-5088

To a mixture of 98 (348 mg, 1.48 mmol), 99 (230 mg, 1.6 mmol) and1,2-dimethoxyethane (7.5 ml), an aqueous 2M sodium carbonate solution(1.5 ml, 3.0 mmol) and tetrakistriphenylphosphine palladium (86 mg,0.074 mmol) were added under an argon atmosphere, and the mixture wasstirred at 90° C. for 6 hours. To the reaction solution, 99 (115 mg,0.816 mmol), an aqueous 2M sodium carbonate solution (0.75 ml, 1.5 mmol)and tetrakistriphenylphosphine palladium (43 mg, 0.037 mmol) were addedunder an argon atmosphere, followed by further stirring at 90° C. for 6hours. The solvent of the reaction solution was distilled off underreduced pressure and the solution was extracted with chloroform. Theextraction liquid was dried and the solvent was distilled off underreduced pressure. The residue was purified by silica gel flash columnchromatography (eluting solvent: n-hexane/ethyl acetate=9/1) and then NHsilica gel flash column chromatography (eluting solvent: n-hexane,n-hexane/ethyl acetate=49/1, 19/1) and then washed with cold n-hexane toobtain THK-5088 (380 mg, 87%) as a yellowish brown solid.

mp 118-120° C., ¹H NMR (500 MHz, DMSO-d₆) δ 1.18 (6H, t, J=7.0 Hz), 3.50(4H, q, J=7.0 Hz), 6.94 (1H, s), 7.32 (1H, dd, J=6.1, 1.5 Hz), 7.43 (1H,d, J=8.4 Hz), 7.89 (1H, d, J=9 Hz), 8.01 (1H, d, J=9 Hz), 8.20 (1H, d,J=8.3 Hz), 8.74 (1H, dt, J=8.3, 2.6 Hz), 9.01 (1H, d, J=2.6 Hz) IR(Nujol) 1465, 1376 cm⁻¹

APCI-MS m/z 296[M+H]⁺

Synthesis Method of THK-5089

Synthesis of 100

To a mixture of 85 (1.00 g, 3.59 mmol), 99 (0.61 g, 4.31 mmol) and1,2-dimethoxyethane (30 ml), potassium carbonate (1.49 g, 10.76 mmol),water (0.62 ml) and tetrakistriphenylphosphine palladium (210 mg, 0.18mmol) were added under an argon atmosphere, and the mixture was stirredat 80° C. for 2.5 hours. The reaction solution was allowed to return toroom temperature and water was added, and the solution was extractedwith ethyl acetate. The extraction liquid was washed with water anddried, and then the solvent was distilled off under reduced pressure.The residue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/9, 1/4) and then recrystallized fromethyl acetate-n-hexane to obtain 100 (1.01 g, 83%) as colorlesscrystals.

mp 189.5-190° C.

APCI-MS m/z 340[M+H]⁺

Synthesis of 101

To an N,N-dimethylformamide (33 ml) solution of 100 (1.00 g, 2.95 mmol),60% sodium hydride (131 mg, 3.28 mmol) was added under ice cooling andstirring under an argon atmosphere, and the mixture was stirred at thesame temperature for 10 minutes and methyl iodide (0.22 ml, 3.54 mmol)was added dropwise at the same temperature, followed by stirring at roomtemperature for 30 minutes. To the reaction solution, ice water wasadded, and the solution was extracted with ethyl acetate. The extractionliquid was washed with water and dried, and then the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/9,1/4) to obtain 101 (1.03 g, 98%) as a colorless solid.

mp 147-148° C.

APCI-MS m/z 354[M+H]⁺

Synthesis of 102

To a chloroform (12 ml) solution of 101 (1.02 g, 2.89 mmol),trifluoroacetic acid (8 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 1.5 hours.To the reaction solution, water was added and the solution was extractedwith ethyl acetate after adjusting the pH to 9 using an aqueouspotassium carbonate solution. The extraction liquid was washed withwater and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: ethyl acetate/n-hexane=1/9) to obtain 102 (732 mg,100%) as a yellow oily substance.

APCI-MS m/z 254[M+H]⁺

Synthesis of THK-5089

To a methanol (32 ml)-acetic acid (3.2 ml) solution, 102 (730 mg, 2.88mmol) and acetoaldehyde (1.63 ml, 28.8 mmol), a picoline borane complex(930 mg, 8.64 mmol) was added little by little under ice cooling andstirring, and the mixture was stirred at room temperature for 16 hours.To the reaction solution, water and an aqueous potassium carbonatesolution were added thereby adjusting the pH to 9, and the solution wasextracted with ethyl acetate. The extraction liquid was washed withwater and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: n-hexane/ethyl acetate=9/1, 4/1) and thenrecrystallized from n-hexane to obtain THK-5089 (680 mg, 84%) as paleyellow crystals.

mp 71-72° C., ¹H NMR (400 MHz, DMSO-d₆) δ 1.20 (3H, t, J=7.1 Hz), 3.04(3H, s), 3.72 (2H, q, J=7.1 Hz), 7.07-7.12 (1H, br), 7.39 (1H, dd,J=8.5, 2.7 Hz), 7.42-7.49 (2H, m), 8.16 (1H, d, J=8.5 Hz), 8.40 (1H, d,J=8.5 Hz), 8.79 (1H, td, J=8.3, 2.7 Hz), 9.09 (1H, d, J=2.7 Hz)

IR (Nujol) 1598 cm⁻¹

APCI-MS m/z 282[M+H]⁺

Synthesis Method of THK-5095

Synthesis of 103

To a 1,2-dimethoxyethane (38 ml) solution of 70 (2.02 g, 4.66 mmol) and66 (1.55 g, 4.66 mmol), potassium carbonate (1.93 g, 13.97 mmol), water(0.81 ml) and tetrakistriphenylphosphine palladium (540 mg, 0.47 mmol)were added under an argon atmosphere, and the mixture was stirred at 80°C. for 16 hours. The reaction solution was allowed to return to roomtemperature and water was added, and the solution was extracted withethyl acetate. The extraction liquid was washed with water and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/4) to obtain 103 (2.82 g, 100%) as apale pink oily substance.

APCI-MS m/z 605[M+H]⁺

Synthesis of 104

To a chloroform (24 ml) solution of 103 (2.81 g, 4.65 mmol),trifluoroacetic acid (16 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 1 hour. Tothe reaction solution, water and ethyl acetate were added, and thesolution was made basic with an aqueous potassium carbonate solution andextracted with ethyl acetate. The extraction liquid was washed withwater and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: n-hexane/ethyl acetate=3/1, 2/1) to obtain 104 (2.28g, 97%) as a pale yellow oily substance.

APCI-MS m/z 505[M+H]⁺

Synthesis of 105

To a chloroform (20 ml) solution of 104 (1.14 g, 2.26 mmol),triethylamine (0.473 ml, 3.39 mmol) and then trifluoroacetic anhydride(0.383 ml, 2.71 mmol) were added dropwise under ice cooling andstirring, and the mixture was stirred at the same temperature for 2hours. To the reaction solution, ice water was added and the solutionwas extracted with ethyl acetate. The extraction liquid was washed withwater and dried and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: ethyl acetate/n-hexane=1/4) to obtain 105 (1.36 g,100%) as a pale yellow oily substance.

APCI-MS m/z 601[M+H]⁺

Synthesis of 106

To a mixture of 105 (1.36 g, 2.26 mmol) and anisole (2 ml),methanesulfonic acid (6 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 20minutes. The reaction solution was ice-cooled and ice water was added,and the solution was made basic with an aqueous potassium carbonatesolution and extracted with ethyl acetate. The extraction liquid waswashed with water and dried and then the solvent was distilled off underreduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/1, ethylacetate) to obtain 106 (890 mg, 94%) as a pale yellow amorphous.

APCI-MS m/z 421[M+H]⁺

Synthesis of 107

To a pyridine (10 ml) solution of 106 (880 mg, 2.09 mmol), atetrahydrofuran (15 ml) solution of paratoluenesulfonic anhydride (1.03g, 3.14 mmol) was added dropwise over 20 minutes under ice cooling andstirring, and the mixture was stirred at the same temperature for 1hour. To the reaction solution, ice water was added, and the solutionwas extracted with ethyl acetate after adjusting the pH to 9 using anaqueous potassium carbonate solution. The extraction liquid was washedwith water and dried the solvent was distilled off under reducedpressure (toluene azeotrope). The residue was purified by silica gelcolumn chromatography (eluting solvent: ethyl acetate/n-hexane=1/2, 1/1,ethyl acetate) to obtain 107 (750 mg, 63%) as a colorless amorphous.

APCI-MS m/z 575[M+H]⁺

Synthesis of THK-5095

To a tetrahydrofuran (10 ml)-water (1 ml) solution of 107 (750 mg, 1.31mmol), lithium hydroxide monohydrate (82 mg, 1.97 mmol) was added underice cooling and stirring, and the mixture was stirred at the sametemperature for 1 hour, followed by stirring at room temperature for 1hour. To the reaction solution, ice water was added and the solution wasextracted with ethyl acetate. The extraction liquid was washed withwater and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: ethyl acetate/n-hexane=1/1, 2/1) and thenrecrystallized from ethyl acetate-n-hexane to obtain THK-5095 (563 mg,90%) as colorless crystals.

mp 109-111° C., ¹H NMR (400 MHz, DMSO-d₆) δ 2.33 (3H, s), 2.76 (3H, d,J=4.5 Hz), 3.56-3.71 (2H, m), 4.30 (1H, dd, J=11, 5.7 Hz), 4.38 (1H, dd,J=11, 3.0 Hz), 4.71-4.76 (1H, m), 5.11 (1H, t, J=5.6 Hz) 6.14 (1H, br),6.66 (2H, d, J=8.8 Hz), 7.01 (1H, dd, J=8.8, 2.4 Hz), 7.31 (1H, d, J=2.1Hz), 7.39 (2H, d, J=8.5 Hz), 7.75 (2H, d, J=8.8 Hz), 7.78 (1H, d, J=9.1Hz), 7.84 (1H, d, J=8.8 Hz), 8.06 (2H, d, J=8.8 Hz), 8.20 (1H, d, J=8.5Hz)

IR (Nujol) 3359, 1741, 1608 cm⁻¹

APCI-MS m/z 479 [M+H]+

Synthesis Method of THK-5096

Synthesis of 108

To a methanol (25 ml)-acetic acid (2.5 ml) solution of 104 (1.13 g, 2.24mmol) and acetoaldehyde (1.26 ml, 22.4 mmol), a picoline borane complex(720 mg, 6.72 mmol) was added under ice cooling and stirring, and themixture was stirred at room temperature for 16 hours. To the reactionsolution, ice water was added, and the solution was extracted with ethylacetate by adjusting the pH to 9 using an aqueous potassium carbonatesolution. The extraction liquid was washed with water and dried, andthen the solvent was distilled off under reduced pressure. The residuewas purified by silica gel column chromatography (eluting solvent:n-hexane/ethyl acetate=9/1, 4/1) to obtain 108 (1.08 g, 91%) as a paleyellow oily substance.

APCI-MS m/z 533 [M+H]⁺

Synthesis of 109

To a mixture of 108 (1.15 g, 2.16 mmol) and anisole (3 ml),methanesulfonic acid (9 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 20minutes. The reaction solution was ice-cooled and ice water was added,and the solution was extracted with ethyl acetate after adjusting the pHto 9 using an aqueous potassium carbonate solution. The extractionliquid was washed with water and dried, and then the solvent wasdistilled off under reduced pressure. The residue was washed with ethylacetate-n-hexane (1/1) to obtain 109 (640 mg, 84%) as yellow crystals.

mp 136-137° C.

APCI-MS m/z 353[M+H]⁺

Synthesis of THK-5096

To a pyridine (10 ml) solution of 109 (630 mg, 1.79 mmol), atetrahydrofuran (10 ml) solution of paratoluenesulfonic anhydride (875mg, 2.68 mmol) was added dropwise over 30 minutes under ice cooling andstirring, and the mixture was stirred at the same temperature for 20minutes. To the reaction solution, ice water was added, and the solutionwas extracted with ethyl acetate after adjusting the pH to 9 using anaqueous potassium carbonate solution. The extraction liquid was washedwith water and dried, and then the solvent was distilled off underreduced pressure (toluene azeotrope). The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/2,ethyl acetate) and then recrystallized from ethyl acetate-n-hexane toobtain THK-5096 (450 mg, 50%) as yellow crystals.

mp 89-90° C., ¹H NMR (400 MHz, DMSO-d₆) δ 1.10 (3H, t, J=7.0 Hz), 2.33(3H, s), 2.96 (3H, s), 3.48 (2H, q, J=7.0 Hz), 3.37-3.71 (2H, m), 4.30(1H, dd, J=11, 5.7 Hz), 4.38 (1H, dd, J=11, 2.7 Hz), 4.71-4.78 (1H, m),5.11 (1H, t, J=5.4 Hz), 6.83 (2H, d, J=9.1 Hz), 7.02 (1H, dd, J=8.9, 2.3Hz), 7.33 (1H, d, J=1.8 Hz), 7.39 (2H, d, J=7.9 Hz), 7.75 (2H, d, J=8.2Hz), 7.79 (1H, d, J=8.8 Hz), 7.87 (1H, d, J=8.8 Hz), 8.11 (2H, d, J=9.1Hz), 8.21 (1H, d, J=9.1 Hz)

IR (Nujol) 1620, 1597 cm⁻¹

APCI-MS m/z 507[M+H]⁺

Synthesis Method of THK-5099

Synthesis of 110

To an N,N-dimethylformamide (30 ml) suspension of 41 (5.84 g, 36.24mmol), thionyl chloride (10.39 ml, 145 mmol) was added dropwise underice cooling and stirring, and the mixture was stirred at the sametemperature for 30 minutes, followed by stirring at room temperature for16 hours. The reaction solution was ice-cooled and ice water was added,and the solution was extracted with ethyl acetate after adjusting the pHto 7 using an aqueous potassium carbonate solution. The extractionliquid was washed with water, dried, purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/9, 1/4) andthen recrystallized from ethyl acetate-n-hexane to obtain 110 (3.88 g,60%) as pale yellow crystals.

mp 176-177° C.

APCI-MS m/z 180[M+H]⁺

Synthesis of 111

To a tetrahydrofuran (30 ml) solution of 110 (700 mg, 3.90 mmol), 22(1.27 g, 4.68 mmol) and triphenylphosphine (1.23 g, 4.68 mmol), atetrahydrofuran (5 ml) solution of diisopropyl azodicarboxylate (0.93ml, 4.68 mmol) was added dropwise under ice cooling and stirring, andthe mixture was stirred at room temperature for 3 days. To the reactionsolution, a tetrahydrofuran (5 ml) solution of 22 (1.27 g, 4.68 mmol)and diisopropyl azodicarboxylate (0.93 ml, 4.68 mmol) were added,followed by further stirring at room temperature for 2 hours. Thesolvent of the reaction solution was distilled off under reducedpressure and the residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/20, 1/15) toobtain 111 (1.18 g, 70%) as a pale yellow oily substance.

APCI-MS m/z 434[M+H]⁺

Synthesis of 112

To a 1,2-dimethoxyethane (25 ml) solution of 111 (1.17 g, 2.70 mmol) and6 (740 mg, 2.70 mmol), potassium carbonate (1.12 g, 8.09 mmol), water(0.5 ml) and tetrakistriphenylphosphine palladium (310 mg, 0.27 mmol)were added under an argon atmosphere, and the mixture was stirred at 80°C. for 7 hours. The reaction solution was allowed to return to roomtemperature and water was added, and the solution was extracted withethyl acetate. The extraction liquid was washed with water and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/9, 1/4) to obtain 112 (1.48 g, 100%)as a yellow oily substance.

APCI-MS m/z 547[M+H]⁺

Synthesis of 113

To a mixture of 112 (1.47 g, 2.69 mmol) and anisole (4 ml),methanesulfonic acid (12 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 1 hour.The reaction solution was poured into ice water, and the solution wasextracted with ethyl acetate after adjusting the pH to 8 using anaqueous potassium carbonate solution. The extraction liquid was washedwith water and dried and then the solvent was distilled off underreduced pressure. The residue was washed with hexane to obtain 113 (900mg, 91%) as pale yellow crystals.

mp 165-166° C.

APCI-MS m/z 367[M+H]⁺

Synthesis of THK-5099

To a pyridine (15 ml) solution of 113 (890 mg, 2.43 mmol), atetrahydrofuran (15 ml) solution of paratoluenesulfonic anhydride (1.19g, 3.64 mmol) was added dropwise over 30 minutes under ice cooling andstirring, and the mixture was stirred at the same temperature for 30minutes. To the reaction solution, ice water was added, and the solutionwas extracted with ethyl acetate after adjusting the pH to 9 using anaqueous potassium carbonate solution. The extraction liquid was washedwith water and dried, and then the solvent was distilled off underreduced pressure (toluene azeotrope). The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/4)and then recrystallized from ethyl acetate to obtain THK-5099 (658 mg,52%) as yellow crystals.

mp 143-143.5° C., ¹HNMR (400 MHz, DMSO-d₆) δ 1.15(6H, t, J=7.0 Hz), 2.31(3H, s), 3.43 (4H, q, J=7.0 Hz), 3.67-3.72 (2H, m), 4.37 (1H, dd, J=11,5.7 Hz), 4.42 (1H, dd, J=11, 3.0 Hz), 4.71-4.77 (1H, m), 5.13 (1H, t,J=5.6 Hz), 6.80 (2H, d, J=9.1 Hz), 6.94 (1H, dd, J=5.6, 3.2 Hz), 7.28(2H, d, J=8.2 Hz), 7.51-7.54 (2H, m), 7.68 (2H, d, J=8.2 Hz), 7.93 (1H,d, J=9.1 Hz), 8.12 (2H, d, J=9.1 Hz), 8.25 (1H, d, J=8.8 Hz)

IR (Nujol) 1593 cm⁻¹

APCI-MS m/z 521[M+H]⁺

Synthesis Method of THK-5105

Synthesis of 114

To an N,N-dimethylformamide (20 ml) suspension of 8 (1.16 g, 10 mmol),thionyl chloride (5 g, 42 mmol) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 16 hours.To the reaction solution, an aqueous saturated sodium hydrogen carbonatesolution was added and the solution was extracted with ethyl acetate.The extraction liquid was washed with water and dried, and then thesolvent was distilled off under reduced pressure. The residue waspurified by silica gel flash column chromatography (eluting solvent:ethyl acetate/n-hexane=1/4) to obtain 114 (1.12 g, 62%) as a colorlesssolid.

mp 190-191° C.

Synthesis of 115

To a tetrahydrofuran (20 ml) solution of 114 (1.0 g, 5.57 mmol), 16 (1.2g, 5.76 mmol) and triphenylphosphine (1.9 g, 7.24 mmol), diisopropylazodicarboxylate (1.46 g, 7.22 mmol) was added dropwise under icecooling and stirring, and the mixture was stirred at room temperaturefor 16 hours. The solvent of the reaction solution was distilled offunder reduced pressure and the residue was purified by silica gel flashcolumn chromatography (eluting solvent: ethyl acetate/n-hexane=1/7) toobtain 115 (1.78 g, 86%) as a pale yellow solid.

mp 50 to 52° C.

Synthesis of 117

To a 1,2-dimethoxyethane (14 ml) solution of 115 (600 mg, 1.62 mmol) and116 (294 mg, 1.78 mmol), potassium carbonate (670 mg, 4.87 mmol), water(0.28 ml) and tetrakistriphenylphosphine palladium (190 mg, 0.16 mmol)were added under an argon atmosphere, and the mixture was stirred at 80°C. for 3 hours. The reaction solution was allowed to return to roomtemperature and water was added, and the solution was extracted withethyl acetate. The extraction liquid was washed with water and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/19, 1/9) to obtain 117 (690 mg, 94%)as a pale yellow solid.

mp 131-133° C.

APCI-MS m/z 455[M+H]⁺

Synthesis of THK-5105

To a tetrahydrofuran (20 ml) solution of 117 (690 mg, 1.52 mmol),tetrabutylammonium fluoride (1.52 ml/1M tetrahydrofuran solution) wasadded, and the mixture was stirred at room temperature for 1 hour. Waterwas added to the reaction solution and the solution was extracted withethyl acetate. The extraction liquid was washed with water and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/4, 1/1) and then recrystallized fromethyl acetate to obtain THK-5105 (437 mg, 85%) as pale yellow crystals.

mp 178-179° C., ¹H NMR (400 MHz, DMSO-d₆) δ 4.07-4.18 (3H, m), 4.44-4.65(2H, m). 5.52 (1H, br), 6.84 (2H, d, J=9.1 Hz), 7.36-7.41 (2H, m), 7.90(1H, d, J=10 Hz), 7.99 (1H, d, J=8.8 Hz), 8.10 (2H, d, J=9.1 Hz), 8.23(1H, d, J=8.8 Hz)

IR (Nujol) 3406, 1616 cm⁻¹

APCI-MS m/z 341[M+H]⁺

Synthesis Method of THK-5106

Synthesis of 118

To a tetrahydrofuran (40 ml) solution of 58 (2.00 g, 10.6 mmol), 16(2.64 g, 12.7 mmol) and triphenylphosphine (3.32 g, 12.7 mmol), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (2.51ml, 12.7 mmol) was added dropwise under ice cooling and stirring, andthe mixture was stirred at the same temperature for 30 minutes, followedby stirring at room temperature for 16 hours. The solvent of thereaction solution was distilled off under reduced pressure and theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/20) to obtain 118 (3.90 g, 100%) as apale yellow oily substance.

APCI-MS m/z 370/372[M+H]⁺

Synthesis of 119

A mixture of 118 (1.83 g, 4.9 mmol), 66 (1.73 g, 5.2 mmol), potassiumcarbonate (1.37 g, 9.9 mmol), tetrakistriphenylphosphine palladium (300mg, 0.26 mmol) and 1,2-dimethoxyethane (45 ml)-water (5 ml) was stirredunder an argon atmosphere at 85° C. for 2 hours andtetrakistriphenylphosphine palladium (283 mg, 0.245 mmol) and potassiumcarbonate (678 mg, 4.9 mmol) were added, followed by stirring at thesame temperature for 4 hours. The reaction solution was allowed toreturn to room temperature and water was added, and the solution wasextracted with ethyl acetate. The extraction liquid was dried and thesolvent was distilled off under reduced pressure and then the residuewas purified by silica gel flash column chromatography (eluting solvent:n-hexane, ethyl acetate/n-hexane=1/9) to obtain 119 (2.28 g, 85%) as apale yellow oily substance.

APCI-MS m/z 541[M+H]⁺

Synthesis of THK-5106

To a chloroform (21 ml) solution of 119 (2.25 g, 3.33 mmol),trifluoroacetic acid (14 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 1 hour andwater (7 ml) was added dropwise, followed by further stirring at roomtemperature for 1 hour. The solvent of the reaction solution wasdistilled off under reduced pressure and chloroform was added, and thesolution was made basic with an aqueous saturated sodium hydrogencarbonate solution and extracted with chloroform/methanol (19/1). Theextraction liquid was dried and the solvent was distilled off underreduced pressure. The residue was purified by silica gel flash columnchromatography (eluting solvent: chloroform, chloroform/methanol=19/1)and then washed with n-hexane and diisopropylether to obtain THK-5106(1.0 g, 92%) as a yellow solid.

mp 137-139° C., ¹H NMR (500 MHz, DMSO-d₆) δ 2.76(3H, d, J=4.4 Hz),4.1-4.2 (3H, m), 4.4-4.7 (2H, m), 5.53 (1H, d, J=4.8 Hz), 6.13 (1H, brs)6.66 (2H, d, J=8.7 Hz), 7.15 (1H, dd, J=9.0, 2.6 Hz), 7.35 (1H, d, J=2.6Hz), 7.80 (1H, d, J=9.0 Hz), 7.83 (1H, d, J=8.7 Hz), 8.06 (2H, d, J=9.0Hz), 8.20 (1H, d, J=8.3 Hz)

IR (Nujol)1612, 1598 cm⁻¹

APCI-MS m/z 327[M+H]⁺

Synthesis Method of THK-5107

Synthesis of 120

To a 1,2-dimethoxyethane (20 ml) solution of 118 (800 mg, 2.16 mmol) and116 (390 mg, 2.38 mmol), potassium carbonate (900 mg, 6.49 mmol), water(0.38 ml) and tetrakistriphenylphosphine palladium (380 mg, 0.22 mmol)were added under an argon atmosphere, and the mixture was stirred 80° C.for 3.5 hours. The reaction solution was allowed to return to roomtemperature and water was added, and the solution was extracted withethyl acetate. The extraction liquid was washed with water and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/19) to obtain 120 (860 mg, 88%) as apale yellow solid.

mp 102-106° C.

APCI-MS m/z 455[M+H]⁺

Synthesis of THK-5107

To a tetrahydrofuran (10 ml) solution of 120 (850 mg, 1.87 mmol),tetrabutylammonium fluoride (1.87 ml/1M tetrahydrofuran solution) wasadded, and the mixture was stirred at room temperature for 30 minutes.Water was added to the reaction solution and the solution was extractedwith ethyl acetate. The extraction liquid was washed with water anddried and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/4, 1/2) to obtain THK-5107 (352 mg,55%) as yellow crystals.

mp 163.5-164° C., ¹H NMR (400 MHz, DMSO-d₆) δ 3.01 (6H, s), 4.09-4.22(3H, m), 4.44-4.65 (2H, m), 5.53 (1H, br), 6.84 (2H, d, J=9.1 Hz), 7.18(1H, dd, J=8.8, 2.4 Hz), 7.38 (1H, d, J=2.4 Hz), 7.83 (1H, d, J=9.1 Hz),7.89 (1H, d, J=8.5 Hz), 8.13 (2H, d, J=9.1 Hz), 8.25 (1H, d, J=8.8 Hz)

IR (Nujol) 1619, 1608, 1595 cm⁻¹

APCI-MS m/z 341[M+H]⁺

Synthesis Method of THK-5111

Synthesis of 121

To a 1,2-dimethoxyethane (65 ml) solution of 59 (2.01 g, 7.99 mmol) and66 (2.66 g, 7.99 mmol), potassium carbonate (3.31 g, 24.0 mmol), water(1.39 ml) and tetrakistriphenylphosphine palladium (920 mg, 0.80 mmol)were added under an argon atmosphere, and the mixture was stirred at 80°C. for 8 hours. The reaction solution was allowed to return to roomtemperature and water was added, and the solution was extracted withethyl acetate. The extraction liquid was washed with water and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/19, 1/9, 1/4) to obtain 121 (3.38 g,100%) as a pale yellow oily substance.

APCI-MS m/z 423[M+H]⁺

Synthesis of 122

To a chloroform (12 ml) solution of 121 (3.37 g, 7.98 mmol),trifluoroacetic acid (8 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 2 hours.To the reaction solution, water was added, and the solution wasextracted with ethyl acetate after adjusting the pH to 9 using anaqueous potassium carbonate solution. The extraction liquid was washedwith water and dried, and then the solvent was distilled off underreduced pressure. The residue was recrystallized from ethyl acetate toobtain 122 (1.71 g, 86%) as yellow crystals.

mp 262-265° C.

APCI-MS m/z 251[M+H]⁺

Synthesis of 123

To a methanol (35 ml)-acetic acid (3.5 ml) solution of 122 (800 mg, 3.20mmol) and acetoaldehyde (1.80 ml, 32.0 mmol), a picoline borane complex(1.03 g, 9.59 mmol) was added under ice cooling and stirring, and themixture was stirred at room temperature for 16 hours. To the reactionsolution, ice water was added, and the solution was extracted with ethylacetate after adjusting the pH to 9 using an aqueous potassium carbonatesolution. The extraction liquid was washed with water and dried, andthen the solvent was distilled off under reduced pressure. The residuewas recrystallized from ethyl acetate to obtain 123 (840 mg, 94%) asyellow crystals.

mp 212-213° C.

APCI-MS m/z 279[M+H]⁺

Synthesis of 124

To a tetrahydrofuran (30 ml) solution of 123 (830 mg, 2.98 mmol), 63(1.29 g, 3.58 mmol) and triphenylphosphine (940 mg, 3.58 mmol), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (0.71ml, 3.58 mmol) was added dropwise over 1 hour under ice cooling andstirring, and the mixture was stirred at the same temperature 1 hour,followed by stirring at room temperature for 16 hours. To the reactionsolution, 63 (540 mg, 1.49 mmol) and triphenylphosphine (390 mg, 1.49mmol) were added under ice cooling and stirring, and diisopropylazodicarboxylate (0.3 ml, 1.49 mmol) and tetrahydrofuran (10 ml) wereadded, followed by further stirring at room temperature for 3 days. Thesolvent of the reaction solution was distilled off under reducedpressure and the residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/9) to obtain124 (2.21 g) as a pale yellow oily substance.

APCI-MS m/z 621[M+H]⁺

Synthesis of 125

To a chloroform (12 ml) solution of 124 (2.21 g), trifluoroacetic acid(8 ml) was added dropwise under ice cooling and stirring and water (2ml) was added, and the mixture was stirred at room temperature for 16hours. To the reaction solution, ice water and ethyl acetate were added,and the solution was extracted with ethyl acetate after adjusting the pHto 9 using an aqueous potassium carbonate solution. The extractionliquid was washed with water and dried, and then the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/2,1/1) to obtain 125 (1.27 g, 84% from 123) as a yellow foam-likesubstance.

APCI-MS m/z 507[M+H]⁺

Synthesis of THK-5111

To a methylene chloride (30 ml) solution of 125 (940 mg, 1.86 mmol),3,4-dihydro-2H-pyran (3.36 ml, 37.2 mmol) and paratoluenesulfonic acidmonohydrate (415 mg, 2.42 mmol) were added, and the mixture was stirredat room temperature for 40 minutes. The reaction solution wasneutralized with triethylamine and the solvent was distilled off underreduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/4) to obtainTHK-5111 (1.05 g, 96%) as a yellow amorphous.

¹H NMR (500 MHz, DMSO-d₆) δ 1.10 (3H, t, J=7.2 Hz), 1.36-1.70 (6H, m),2.34 (3H, s). 2.97 (3H, s), 3.37-3.45 (1H, m), 3.49 (2H, q, J=7.2 Hz),3.68-3.74, 3.84-3.90 (1H, m), 4.13-4.37 (5H, m), 4.71-4.74, 4.86-4.88(1H, m), 6.84 (2H, d, J=8.7 Hz), 7.06 (1H, d, J=8.7 Hz), 7.31 (1H, br),7.39-7.44 (2H, m), 7.78-7.84 (3H, m), 7.90 (1H, d, J=9.0 Hz), 8.12 (2H,d, J=9.0 Hz), 8.27 (1H, br)

APCI-MS m/z 591[M+H]⁺

Synthesis Method of THK-5112

Synthesis of THK-5112

To a methanol (10 ml)-acetic acid (1 ml) solution of THK-5106 (478 mg,1.46 mmol) and acetoaldehyde (323 mg, 7.3 mmol), a picoline boranecomplex (204 mg, 1.9 mmol) was added under ice cooling and stirring andthe mixture was stirred at room temperature for 1 hour, andacetoaldehyde (323 mg, 7.3 mmol) and a picoline borane complex (204 mg,1.9 mmol) were added, followed by further stirring at room temperaturefor 5 hours. The solvent of the reaction solution was distilled offunder reduced pressure and aqueous 10% hydrochloric acid (5 ml) wasadded to the residue. After stirring at room temperature for 30 minute,the solution was made basic by adding potassium carbonate. The reactionsolution was extracted with chloroform and the extraction liquid wasdried, and then the solvent was distilled off under reduced pressure.The residue was purified by NH silica gel flash column chromatography(eluting solvent: n-hexane, n-hexane/ethyl acetate=1/1) to obtain as ayellow oily substance. After dissolving by adding oxalic acid (292 mg,2.9 mmol) to an acetone (10 ml) solution of the present product, thesolvent was distilled off under reduced pressure. The residue was washedwith diisopropylether and diethyl ether to obtain THK-5112 (660 mg, 91%)as an orange solid.

mp 193-194.5° C. (dec), ¹H NMR (400 MHz, DMSO-d₆) δ 1.10 (3H, t, J=7.0Hz), 2.97 (3H, s), 3.49 (2H, q, J=7.0 Hz), 4.0-4.2 (3H, m), 4.4-4.7 (2H,m), 6.83 (2H, d, J=9.1 Hz), 7.18 (1H, dd, J=8.7, 2.4 Hz), 7.38 (1H, d,J=2.4 Hz), 7.84 (1H, d, J=9.1 Hz), 7.89 (1H, d, J=8.7 Hz), 8.12 (2H, d,J=9.0 Hz), 8.27 (1H, d, J=8.8 Hz)

IR (Nujol) 2922, 2852 cm⁻¹

APCI-MS m/z 355[M+H]⁺

Synthesis Method of THK-5113

Synthesis of 126

To a methylene chloride (20 ml) suspension of 122 (900 mg, 3.60 mmol)and triethylamine (1.51 ml, 10.79 mmol), trifluoro acid anhydride (1.22ml, 8.63 mmol) was added dropwise under ice cooling and stirring, andthe mixture was stirred at the same temperature for 30 minutes. To thereaction solution, ice water was added and the solution was extractedwith chloroform/methanol. The extraction liquid was dried and thesolvent was distilled off under reduced pressure, and then the residuewas recrystallized from ethyl acetate-n-hexane to obtain 126 (1.24 g,99%) as orange crystals.

mp 176-178° C.

APCI-MS m/z 347[M+H]⁺

Synthesis of 127

To a tetrahydrofuran (30 ml) suspension of 126 (1.23 g, 3.55 mmol), 63(1.54 g, 4.26 mmol) and triphenylphosphine (1.12 g, 4.26 mmol), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (0.84ml, 4.26 mmol) was added dropwise over 20 minutes under ice cooling andstirring, and the mixture was stirred at the same temperature 2 hours,followed by stirring at room temperature for 16 hours. To the reactionsolution, 63 (640 mg, 1.78 mmol), triphenylphosphine (470 mg, 1.78 mmol)and diisopropyl azodicarboxylate (0.35 ml, 1.78 mmol) were added,followed by stirring at room temperature for 4 hours. Furthermore, 63(640 mg, 1.78 mmol), triphenylphosphine (470 mg, 1.78 mmol) anddiisopropyl azodicarboxylate (0.35 ml, 1.78 mmol) were added to thereaction solution, followed by stirring at room temperature for 16hours. The solvent of the reaction solution was distilled off underreduced pressure and the residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/4) to obtain127 (3.28 g) as a pale yellow oily substance.

APCI-MS m/z 347[M+H]⁺

Synthesis of 128

To a chloroform (12 ml) solution of 127 (3.27 g), trifluoroacetic acid(8 ml) was added dropwise and water (2 ml) was added under ice coolingand stirring, and the mixture was stirred at room temperature for 8hours. To the reaction solution, ice water and ethyl acetate were added,and the solution was extracted with ethyl acetate after adjusting the pHto 7 using an aqueous potassium carbonate solution. The extractionliquid was washed with water and dried, and then the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/2,1/1) to obtain 128 (1.85 g, 91% from 126) as a pale yellow foam-likesubstance.

APCI-MS m/z 575[M+H]⁺

Synthesis of 129

To a tetrahydrofuran (60 ml) solution of 128 (1.84 g, 3.20 mmol) and3,4-dihydro-2H-pyran (5.81 ml, 64.1 mmol), paratoluenesulfonic acidmonohydrate (720 mg, 4.16 mmol) was added, and the mixture was stirredat room temperature for 16 hours. The reaction solution was neutralizedwith triethylamine and the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: ethyl acetate/n-hexane=1/9, 1/4, 1/2) to obtain 129(2.10 g, 99%) as a pale yellow foam-like substance.

APCI-MS m/z 659[M+H]⁺

Synthesis of THK-5113

To a tetrahydrofuran (25 ml)-water (10 ml) solution of 129 (2.09 g, 3.17mmol), lithium hydroxide monohydrate (200 mg, 4.76 mmol) was added underice cooling and stirring, and the mixture was stirred at the sametemperature for 30 minutes. The reaction solution was extracted withethyl acetate, and the extraction liquid was dried and the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/2)to obtain THK-5113 (1.70 g, 95%) as a pale yellow amorphous.

¹H NMR (400 MHz, DMSO-d₆) δ 1.36-1.72 (6H, m), 2.34 (3H, s), 2.76 (3H,d, J=4.2 Hz), 3.35-3.48, 3.67-3.76, 3.83-3.91 (2H, m), 4.11-4.37 (5H,m), 4.70-4.74, 4.85-4.88 (1H, m), 6.14 (1H, br), 6.66 (2H, d, J=8.8 Hz),7.03 (1H, dd, J=8.8, 2.4 Hz), 7.24-7.28 (1H, m), 7.39-7.44 (2H, m),7.76-7.87 (4H, m), 8.07 (2H, d, J=8.8 Hz), 8.21 (1H, d, J=8.8 Hz)

IR (Nujol) 3422, 1609, 1596 cm⁻¹

APCI-MS m/z 563[M+H]⁺

Synthesis Method of THK-5115

Synthesis of THK-5115

To a methanol (20 ml)-acetic acid (2 ml) solution of THK-5113 (1.02 g,1.81 mmol) and an aqueous 37% formalin solution (1.45 ml, 18.1 mmol), apicoline borane complex (580 mg, 5.44 mmol) was added under ice coolingand stirring, and the mixture was stirred at the same temperature for 5minutes, followed by stirring at room temperature for 1 hour. To thereaction solution, ice water was added, and the solution was extractedwith ethyl acetate after adjusting the pH to 9 using an aqueouspotassium carbonate solution. The extraction liquid was washed withwater and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: n-hexane/ethyl acetate=4/1) to obtain THK-5115 (980mg, 93%) as a yellow amorphous. ¹H NMR (500 MHz, DMSO-d₆) δ 1.36-1.71(6H, m), 2.34 (3H, s), 3.01 (6H, s), 3.37-3.47 (1H, m), 3.69-3.74,3.84-3.90 (1H, m), 4.13-4.37 (5H, m), 4.71-4.74, 4.85-4.88 (1H, m), 6.85(2H, d, J=9.0 Hz), 7.05 (1H, d, J=8.7 Hz), 7.30 (1H, br), 7.40-7.44 (2H,m), 7.78-7.84 (3H, m), 7.90 (1H, d, J=9.3 Hz), 8.14 (2H, d, J=8.7 Hz),8.25 (1H, br)

IR (Nujol) 1731, 1620, 1595 cm⁻¹

APCI-MS m/z 577[M+H]⁺

Synthesis Method of THK-5116

Synthesis of 131

A mixture of 115 (625 mg, 1.69 mmol), 130 (593 mg, 1.86 mmol), potassiumcarbonate (467 mg, 3.40 mmol), tetrakistriphenylphosphine palladium (195mg, 0.169 mmol) and 1,2-dimethoxyethane (4.5 ml)-water (0.5 ml) wasstirred under an argon atmosphere at 85° C. for 16 hours, andtetrakistriphenylphosphine palladium (98 mg, 0.085 mmol) was added,followed by further stirring at the same temperature for 8 hours. Thereaction solution was allowed to return to room temperature andchloroform was added, and then insolubles were removed by filtration andthe solution was washed with chloroform. The chloroform layers werecombined and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel flash columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/19, 1/9, 1/6)to obtain 131 (799 mg, 90%) as a pale brown solid.

APCI-MS m/z 527[M+H]⁺

Synthesis of THK-5116

To a chloroform (10 ml) solution of 131 (790 mg, 1.50 mmol),trifluoroacetic acid (5 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 1 hour andwater (5 ml) was added dropwise, followed by further stirring at roomtemperature for 1 hour. After the organic the solvent was distilled offunder reduced pressure, the reaction solution was made basic under icecooling using an aqueous saturated potassium carbonate solution. Theprecipitate was collected by filtration, washed with water and thendried. The obtained solid was purified by NH silica gel flash columnchromatography (eluting solvent: chloroform, chloroform/methanol=9/1,1/1) and then washed with methanol to obtain THK-5116 (396 mg, 84%) as ayellow solid. mp 219-221° C., ¹H NMR (500 MHz, DMSO-d₆)δ4.0-4.2 (3H, m),4.4-4.7 (2H, m), 5.53 (3H, brs), 6.67 (2H, d, J=8.6 Hz), 7.3-7.4 (2H,m), 7.87 (1H, d, J=7.3 Hz), 7.92 (1H, d, J=8.6 Hz), 7.95 (2H, d, J=8.7Hz), 8.19 (1H, d, J=8.6 Hz)

IR (Nujol) 1623, 1600 cm⁻¹

APCI-MS m/z 313[M+H]⁺

Synthesis Method of THK-5117

Synthesis of 132

A mixture of 115 (625 mg, 1.69 mmol), 66 (619 mg, 1.86 mmol), potassiumcarbonate (467 mg, 3.4 mmol), tetrakistriphenylphosphine palladium (195mg, 0.169 mmol) and 1,2-dimethoxyethane (4.5 ml)-water (0.5 ml) wasstirred under an argon atmosphere at 85° C. for 16 hours. The reactionsolution was allowed to return to room temperature and water was added,and the solution was extracted with ethyl acetate. The extraction liquidwas dried and the solvent was distilled off under reduced pressure, andthen the residue was purified by silica gel flash column chromatography(eluting solvent: n-hexane, ethyl acetate/n-hexane=1/19, 1/9, 1/6) toobtain 132 (920 mg, 100%) as a colorless oily substance.

APCI-MS m/z 541[M+H]⁺

Synthesis of THK-5117

To a chloroform (10 ml) solution of 132 (912 mg, 1.69 mmol),trifluoroacetic acid (5 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 1 hour andwater (5 ml) was added dropwise, followed by further stirring at roomtemperature for 1 hour. After the organic the solvent was distilled offunder reduced pressure, the reaction solution was made basic under icecooling using an aqueous saturated potassium carbonate solution, and thesolution was extracted with chloroform-methanol. The extraction liquidwas dried and the solvent was distilled off under reduced pressure, andthen the residue was washed with diisopropylether to obtain THK-5117(468 mg, 85%) as a yellow solid.

mp 158-159° C., ¹H NMR (500 MHz, DMSO-d₆) δ 2.75(3H, d, J=1.8 Hz),4.0-4.2 (3H, m), 4.4-4.7 (2H, m), 5.53 (1H, d, J=4.8 Hz), 6.09 (1H,brs), 6.65 (2H, d, J=8.9 Hz), 7.3-7.4 (2H, m), 7.87 (1H, d, J=8.7 Hz),7.94 (1H, d, J=8.7 Hz), 8.03 (2H, d, J=8.6 Hz), 8.20 (1H, d, J=8.7 Hz)

IR (Nujol) 1600 cm⁻¹

APCI-MS m/z 327[M+H]⁺

Synthesis Method of THK-5100

Synthesis of 133

To a N,N-dimethylformamide (10 ml) solution of 93 (790 mg, 2.34 mmol),60% sodium hydride (112 mg, 2.81 mmol) was added under ice cooling andstirring under an argon atmosphere, and the mixture was stirred at thesame temperature for 5 minutes and methyl iodide (0.22 ml, 3.5 mmol) wasadded dropwise, followed by stirring at the same temperature for 10minutes. To the reaction solution, ice water was added, and the solutionwas extracted with ethyl acetate. The extraction liquid was washed withwater and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: ethyl acetate/n-hexane=1/9, 1/4) to obtain 133 (800mg, 97%) as a pale yellow solid.

mp 144-145° C.

APCI-MS m/z 353[M+H]⁺

Synthesis of 134

To a chloroform (12 ml) solution of 133 (790 mg, 2.24 mmol),trifluoroacetic acid (8 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 1 hour. Tothe reaction solution, ice water was added, and the solution wasextracted with ethyl acetate after adjusting the pH to 9 using anaqueous potassium carbonate solution. The extraction liquid was washedwith water and dried, and then the solvent was distilled off underreduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/4) to obtain134 (568 mg, 100%) as a pale yellow solid.

mp 134-134.5° C.

APCI-MS m/z 253[M+H]⁺

Synthesis of THK-5100

To a methanol (24 ml)-acetic acid (2.4 ml) solution of 134 (560 mg, 2.22mmol) and acetoaldehyde (1.25 ml, 22.2 mmol), a picoline borane complex(710 mg, 6.66 mmol) was added little by little under ice cooling andstirring, and the mixture was stirred at room temperature for 16 hours.To the reaction solution, ethyl acetate was added, and the solution wasextracted with ethyl acetate after adjusting the pH to 9 using anaqueous potassium carbonate solution. The extraction liquid was washedwith water and dried, and then the solvent was distilled off underreduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: n-hexane/ethyl acetate=9/1) and thenwashed with n-hexane to obtain THK-5100 (475 mg, 76%) as pale yellowcrystals.

mp 137-138° C., ¹H NMR (400 MHz, DMSO-d₆) δ 1.11 (3H, t, J=7.1 Hz), 3.01(3H, s), 3.55 (2H, q, J=7.1 Hz), 6.93 (1H, d, J=2.7 Hz), 7.33 (2H, t,J=8.9 Hz), 7.46 (1H, dd, J=9.4, 3.0 Hz), 7.87 (1H, d, J=9.4 Hz), 7.95(1H, d, J=8.8 Hz), 8.15 (1H, d, J=8.8 Hz), 8.24 (2H, dd, J=9.1, 5.7 Hz)

IR (Nujol) 1618 cm⁻¹

APCI-MS m/z 281[M+H]⁺

Synthesis Method of THK-5091 and THK-5092

Synthesis of 135

To a mixture of 79 (627 mg, 3.0 mmol), 99 (465 mg, 3.3 mmol) and1,2-dimethoxyethane (20 ml), an aqueous 2M sodium carbonate solution (3ml, 6.0 mmol) and tetrakistriphenylphosphine palladium (173 mg, 0.15mmol) were added under an argon atmosphere, and the mixture was heatedat reflux for 3 hours. The reaction solution was allowed to return toroom temperature, diluted with ethyl acetate and then washed in turnwith water and saturated saline. The organic layer was dried and thesolvent was distilled off under reduced pressure, and then the residuewas purified by silica gel flash column chromatography (eluting solvent:n-hexane/ethyl acetate=4/1, 3/1) to obtain 135 (700 mg, 87%) as a paleyellow solid.

mp 159-161° C.

APCI-MS m/z 270 [M+H]⁺

Synthesis of 136

A mixture of 135 (260 mg, 0.96 mmol), 10% Pd—C (100 mg), ethanol (30 ml)and ethyl acetate (10 ml) was stirred under a hydrogen atmosphere atroom temperature for 16 hours. The catalyst was removed by filtrationand the solvent of the filtrate was distilled off under reducedpressure, and then the residue was washed with diisopropylether toobtain 136 (178 mg, 77%) as a solid.

mp 156-157° C.

APCI-MS m/z 240[M+H]⁺

Synthesis of THK-5091, THK-5092

To a methanol (20 ml)-acetic acid (1 ml) solution of 136 (170 mg, 0.71mmol) and acetoaldehyde (300 mg, 7 mmol), a picoline borane complex (230mg, 2.14 mmol) was added little by little under ice cooling andstirring, and the mixture was stirred at room temperature for 8 hours.To the reaction solution, saturated saline was added and the solutionwas extracted with ethyl acetate.

The extraction liquid was washed with saturated saline and dried, andthen the solvent was distilled off under reduced pressure. The residuewas purified by silica gel column chromatography (eluting solvent:n-hexane/ethyl acetate=10/1) to obtain THK-5092 (120 mg, 57%) andTHK-5091 (70 mg, 41%) in the order of elution.

THK-5092

Yellow oily substance, ¹H NMR (400 MHz, CDCl₃) δ 1.07 (6H, t, J=7.2 Hz),3.22 (4H, q, J=7.2 Hz), 7.09 (1H, dd, J=8.2, 2.4 Hz), 7.20 (1H, dd,J=7.3, 0.9 Hz), 7.67 (1H, dd, J=9.6, 8.6 Hz), 7.80 (1H, d, J=9.1 Hz),7.85 (1H, d, J=8.5 Hz), 8.66 (1H, m), 8.72 (1H, dd, J=8.8, 0.9 Hz), 8.94(1H, d, J=2.7 Hz)

IR (Nujol) 1586 cm⁻¹

APCI-MS m/z 296[M+H]⁺

THK-5091

mp 175-176° C., ¹H NMR (400 MHz, CDCl₃) δ 1.48 (3H, t, J=7.3 Hz), 3.35(2H, q, J=7.3 Hz), 4.27 (1H, brs), 6.66 (1H, d, J=7.0 Hz), 7.08 (1H, dd,J=7.9, 2.1 Hz), 7.52 (1H, d, J=8.5 Hz), 7.61 (1H, t), 7.75 (1H, d, J=8.8Hz), 8.27 (1H, d, J=8.8 Hz), 8.66 (1H, m), 8.93 (1H, d, J=2.7 Hz)

IR (Nujol) 3332, 1610 cm⁻¹

APCI-MS m/z 268[M+H]⁺

Synthesis Method of THK-5097

Synthesis of 137

A mixture of 15 (3.73 g, 23 mmol), triisopropylsilane chloride (5.0 g,26 mmol), imidazole (2.07 g, 26 mmol) and N,N-dimethylformamide (50 ml)was stirred at room temperature for 16 hours. To the reaction solution,ice water was added and the solution was extracted with ethyl acetate.The extraction liquid was washed with water and dried, and then thesolvent was distilled off under reduced pressure. The residue waspurified by silica gel flash column chromatography (eluting solvent:n-hexane, n-hexane/ethyl acetate=3/1, 1/1) and then washed with n-hexaneto obtain 137 (6.52 g, 89%) as a pale brown solid.

Synthesis of 138

To a methylene chloride (40 ml) solution of 137 (6.36 g, 20 mmol) and 4(5.76 g, 28 mmol), a methylene chloride (10 ml) solution oftrifluoromethanesulfonic anhydride (4.3 ml, 26 mmol) was added dropwiseunder ice cooling and stirring, and the mixture was stirred at the sametemperature 1 hour. To the reaction solution, ice water was added andinsolubles were removed by filtration, followed by washing withchloroform. The organic layers were combined and dried, and then thesolvent was distilled off under reduced pressure. The residue waspurified by silica gel flash column chromatography (eluting solvent:n-hexane, n-hexane/ethyl acetate=19/1, 9/1) to obtain 138 (8.60 g, 95%)as a brown oily substance.

Synthesis of 139

To a mixture of 138 (4.24 g, 9.4 mmol), 99 (1.54 g, 10.9 mmol) and1,2-dimethoxyethane (80 ml), tripotassium phosphate (3.98 g) andtetrakistriphenylphosphine palladium (575 mg, 0.498 mmol) were addedunder an argon atmosphere, and the mixture was heated at reflux for 16hours. The reaction solution was allowed to return to room temperatureand insolubles were removed by filtration, followed by washing withchloroform. The organic layers were combined and the solvent wasdistilled off under reduced pressure. To the residue, chloroform-waterwas added and the solution was extracted with chloroform. The extractionliquid was dried and the solvent was distilled off under reducedpressure, and then the residue was purified by silica gel flash columnchromatography (eluting solvent: n-hexane, n-hexane/ethyl acetate=50/1,20/1) to obtain 139 (3.31 g, 88%) as a pale brown oily substance.

Synthesis of 140

To 139 (3.31 g, 8.35 mmol), tetrahydrofuran (4 ml) and thentetrabutylammonium fluoride (8.5 ml/1M tetrahydrofuran solution) wereadded, and the mixture was stirred at room temperature for 1 hour. Thesolvent of the reaction solution was distilled off under reducedpressure, and the residue was dissolved in chloroform and washed withwater. The organic layer was dried and then the solvent was distilledoff under reduced pressure. The residue was purified by silica gel flashcolumn chromatography (eluting solvent: chloroform,chloroform/methanol=50/1, 20/1, 10/1) and then washed with n-hexane anddiisopropylether to obtain 140 (1.87 g, 93%) as a pale brown solid.

Synthesis of 141

To a methylene chloride (20 ml) solution of 140 (1.87 g, 7.8 mmol) andethyldiisopropylamine (2.6 ml, 15.3 mmol), a methylene chloride (10 ml)solution of trifluoromethanesulfonic anhydride (2.35 ml, 14.3 mmol) wasadded dropwise under ice cooling and stirring, and the mixture wasstirred at room temperature for 1.5 hours. The reaction solution waspoured into ice water and the solution was extracted with chloroform.The extraction liquid was dried and the solvent was distilled off underreduced pressure, and then residue was purified by silica gel flashcolumn chromatography (eluting solvent: n-hexane, n-hexane/ethylacetate=50/1, 20/1, 10/1) to obtain 141 (1.81 g, 62%) as a pale yellowsolid.

Synthesis of 142

To a mixture of 141 (2.03 g, 5.3 mmol), acetoamide (393 mg, 6.65 mmol)and 1,4-dioxane (50 ml), cesium carbonante (2.49 g), Pd₂(dba)₃ (251 mg)and Xantphos (316 mg) were added under an argon atmosphere, and themixture was heated at reflux for 16 hours. The reaction solution wasallowed to return to room temperature and insolubles were removed byfiltration, followed by washing with chloroform. The organic layers werecombined and the solvent was distilled off under reduced pressure. Tothe residue, chloroform-water was added and the solution was extractedwith chloroform. The extraction liquid was dried and the solvent wasdistilled off under reduced pressure, and then the residue was purifiedby silica gel flash column chromatography (eluting solvent: chloroform,chloroform/methanol=50/1, 20/1) to obtain 142 (1.35 g, 88%) as a palebrown solid.

Synthesis of 143

To an N,N-dimethylformamide (8 ml) solution of 142 (650 mg, 2.3 mmol),60% sodium hydride (111 mg, 2.7 mmol) was added under ice cooling andstirring under an argon atmosphere, and the mixture was stirred at thesame temperature for 20 minutes and an N,N-dimethylformamide (4 ml)solution of ethyl iodide (396 mg, 2.5 mmol) was added dropwise at thesame temperature, followed by stirring at room temperature for 1 hour.To the reaction solution, ice water was added and the solution wasextracted with ethyl acetate. The extraction liquid was washed withwater and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel flash columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/2, 1/1, 2/1)and then washed with n-hexane to obtain 143 (575 mg, 80%) as a palebrown solid.

APCI-MS m/z 310[M+H]⁺

Synthesis of 144

To a tetrahydrofuran (2 ml) solution of 143 (531 mg, 1.7 mmol), Zr(BH₄)₂(14 ml/0.24M tetrahydrofuran solution) was added dropwise under icecooling and stirring, and the mixture was stirred at the sametemperature for 30 minutes. To the reaction solution, methanol was addedand the solvent was distilled off under reduced pressure. To theresidue, an aqueous saturated sodium hydrogen carbonatesolution-chloroform was added and insolubles were removed by filtration,and then the aqueous layer was extracted with chloroform. The extractionliquid was dried and the solvent was distilled off under reducedpressure, and then the residue was purified by silica gel flash columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/20, 1/10) toobtain 144 (249 mg, 49%) as a yellow oily substance.

APCI-MS m/z 296[M+H]⁺

Synthesis of THK-5097

After dissolving by adding oxalic acid (150 mg, 1.7 mmol) to an acetone(10 ml) solution of 144 (246 mg, 0.83 mmol), the solvent was distilledoff under reduced pressure. The residue was washed with diisopropyletherto obtain THK-5097 (305 mg, 84%) as an orange solid.

mp 127-129° C., ¹H NMR (400 MHz, DMSO-d₆) δ 1.19 (6H, t, J=7.0 Hz), 3.52(4H, q, J=7.0 Hz), 7.01 (1H, d, J=2.5 Hz), 7.27 (1H, d, J=2.5 Hz), 7.34(1H, dd, J=8.6, 2.5 Hz), 7.7-7.8 (2H, m), 8.24 (1H, d, J=8.5 Hz),8.7-8.8 (1H, m), 9.02 (1H, d, J=9.7 Hz)

IR (Nujol) 1659 cm⁻¹

APCI-MS m/z 296[M+H]⁺

Synthesis of THK-5098

Synthesis of 145

To an N,N-dimethylformamide (6 ml) solution of 142 (650 mg, 2.3 mmol),60% sodium hydride (111 mg, 2.7 mmol) was added under ice cooling andstirring under an argon atmosphere, and the mixture was stirred at thesame temperature for 20 minutes and an N,N-dimethylformamide (4 ml)solution of ethyl iodide (360 mg, 2.5 mmol) was added dropwise at thesame temperature, followed by stirring at room temperature for 1 hour.To the reaction solution, ice water was added and the solution wasextracted with ethyl acetate. The extraction liquid was dried and thesolvent was distilled off under reduced pressure. The residue waspurified by silica gel flash column chromatography (eluting solvent:ethyl acetate/n-hexane=1/2, 1/1, 2/1) and then washed with n-hexane toobtain 145 (518 mg, 76%) as a pale brown solid.

Synthesis of 146

To a tetrahydrofuran (4 ml) solution of 145 (517 mg, 1.75 mmol),Zr(BH₄)₂ (14 ml/0.24M tetrahydrofuran solution) was added dropwise underice cooling and stirring, and the mixture was stirred at roomtemperature for 30 minutes. The reaction solution was ice-cooled andmethanol was added, and the solvent was distilled off under reducedpressure. To the residue, methanol was added and methanol was distilledoff under reduced pressure. To the residue, an aqueous saturated sodiumhydrogen carbonate solution-ethyl acetate was added and insolubles wereremoved by filtration, and then aqueous layer was extracted with ethylacetate. The extraction liquid was dried and the solvent was distilledoff under reduced pressure, and then the residue was purified by silicagel flash column chromatography (eluting solvent: ethylacetate/n-hexane=1/20, 1/10) to obtain 146 (258 mg, 52%) as a yellowoily substance.

Synthesis of THK-5098

After dissolving by adding oxalic acid (164 mg, 1.8 mmol) to an acetone(10 ml) solution of 146 (257 mg, 0.91 mmol), the solvent was distilledoff under reduced pressure. The residue was washed with diisopropyletherto obtain THK-5098 (333 mg, 88%) as an orange solid.

¹H NMR (400 MHz, DMSO-d₆) δ 1.13 (3H, t, J=7.0 Hz), 3.05 (3H, s), 3.58(2H, q, J=7.0 Hz), 7.03 (1H, d, J=2.5 Hz), 7.3-7.4 (2H, m), 7.8-7.9 (2H,m), 8.25 (1H, d, J=8.1 Hz), 8.7-8.8 (1H, m), 9.03 (1H, d, J=2.5 Hz)

IR (Nujol) 1646 cm⁻¹

APCI-MS m/z 282[M+H]⁺

Synthesis Method of THK-5119

Synthesis of 147

To a tetrahydrofuran (15 ml) solution of 114 (840 mg, 4.68 mmol) andethyl vinyl ether (1.35 ml, 10.0 mmol), paratoluenesulfonic acidpyridine salt (120 mg, 0.47 mmol) was added, and the mixture was stirredat room temperature for 2.5 hours. After adjusting the pH of thereaction solution to 9 by adding triethylamine, the solvent wasdistilled off under reduced pressure and the residue was purified bysilica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/9) to obtain 147 (970 mg, 82%) as a colorless oilysubstance.

APCI-MS m/z 252/254[M+H]⁺

Synthesis of 148

To a 1,2-dimethoxyethane (70 ml) solution of 147 (2.08 g, 8.26 mmol) and6a (2.90 g, 9.09 mmol), potassium carbonate (3.43 g, 24.8 mmol), water(1.44 ml) and tetrakistriphenylphosphine palladium (960 mg, 0.83 mmol)were added under an argon atmosphere, and the mixture was stirred at 80°C. for 9 hours. The reaction solution was allowed to return to roomtemperature and water was added, and the solution was extracted withethyl acetate. The extraction liquid was washed with water and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/9) to obtain 148 (3.00 g, 89%) as acolorless solid.

mp 115-117° C.

APCI-MS m/z 409[M+H]⁺

Synthesis of 149

To an N,N-dimethylformamide (25 ml) solution of 148 (1.79 g, 4.38 mmol),60% sodium hydride (210 mg, 5.25 mmol) was added under ice cooling andstirring and an argon atmosphere at the same temperature. After stirringat the same temperature for 10 minutes, methyl iodide (0.41 ml, 6.57mmol) was added dropwise and the mixture was stirred at the sametemperature for 10 minutes. To the reaction solution, ice water wasadded and the solution was extracted with ethyl acetate. The extractionliquid was washed with water and dried, and then the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/4)to obtain 149 (1.85 g, 100%) as a colorless resinous substance.

APCI-MS m/z 423[M+H]⁺

Synthesis of 150

To a chloroform (12 ml) solution of 149 (1.85 g, 4.38 mmol),trifluoroacetic acid (8 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 1 hour. Tothe reaction solution, ice water was added and the solution wasextracted with ethyl acetate. The extraction liquid was washed withwater and dried, and then the solvent was distilled off under reducedpressure. The residue was ground and then washed with ethylacetate/n-hexane (1/4) to obtain 150 (1.04 g, 95%) as orange crystals.

mp 273-275° C.

APCI-MS m/z 251[M+H]⁺

Synthesis of 151

To a methylene chloride (40 ml) suspension of 150 (1.03 g, 4.12 mmol)and triethylamine (1.72 ml, 12.36 mmol), trifluoroacetic anhydride (1.40ml, 9.89 mmol) was added dropwise under ice cooling and stirring, andthe mixture was stirred at the same temperature for 30 minutes. To thereaction solution, ice water was added and the solution was extractedwith ethyl acetate. The extraction liquid was dried and the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/2)and then purified by NH silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/1, ethyl acetate) to obtain 151 (1.40g, 98%) as a pale yellow solid.

mp 214-216° C.

APCI-MS m/z 347[M+H]⁺

Synthesis of 152

To a tetrahydrofuran (40 ml) suspension of 151 (1.39 g, 4.01 mmol), 63(1.74 g, 4.82 mmol) and triphenylphosphine (1.26 g, 4.82 mmol), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (0.95ml, 4.82 mmol) added dropwise over 30 minutes under ice cooling andstirring, and the mixture was stirred at the same temperature for 1hour, followed by stirring at room temperature for 16 hours. To thereaction solution, 63 (1.45 g, 4.01 mmol), triphenylphosphine (1.05 g,4.01 mmol) and diisopropyl azodicarboxylate (0.80 ml, 4.01 mmol) wereadded, followed by further stirring at room temperature for 1 hour. Thesolvent of the reaction solution was distilled off under reducedpressure and the residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/4) to obtain acolorless oily substance (5.65 g). To a chloroform (24 ml) solution ofthe present product (5.65 g), trifluoroacetic acid (16 ml) was addeddropwise under ice cooling and stirring and water was added (16 ml),followed by stirring at room temperature for 3 days. The reactionsolution was extracted with ethyl acetate after adjusting the pH to 9using an aqueous potassium carbonate solution. The extraction liquid wasdried and the solvent was distilled off under reduced pressure and theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/4, 2/3) to obtain 152 (2.15 g, 93%) asa yellow amorphous.

APCI-MS m/z 575[M+H]⁺

Synthesis of 153

To a methylene chloride (60 ml) solution of 152 (2.14 g, 3.72 mmol) and3,4-dihydro-2H-pyran (6.75 ml, 74 mmol), paratoluenesulfonic acidmonohydrate (830 mg, 4.84 mmol) was added, and the mixture was stirredat room temperature for 1 hour. After adjusting the pH of the reactionsolution to 9 suing triethylamine, the solvent was distilled off underreduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/9, 1/2) toobtain 153 (2.36 g, 96%) as a pale yellow amorphous.

APCI-MS m/z 659 [M+H]+

Synthesis of THK-5119

To a tetrahydrofuran (28 ml)-water (11 ml) solution of 153 (2.35 g, 3.57mmol), lithium hydroxide monohydrate (225 mg, 5.35 mmol) was added underice cooling and stirring, and the mixture was stirred at the sametemperature for 30 minutes. Water was added to the reaction solution andthe solution was extracted with ethyl acetate. The extraction liquid waswashed with water and dried, and then the solvent was distilled offunder reduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=2/3) to obtainTHK-5119 (1.78 g, 89%) as pale yellow crystals.

mp 126-126.5° C.

¹H NMR (500 MHz, DMSO-d₆) δ 1.35-1.71 (6H, m), 2.34, 2.34 (3H, s), 2.75(3H, br), 3.38-3.46 (1H, m), 3.66-3.72, 3.81-3.88 (1H, m), 4.08-4.37(5H, m), 4.69-4.72, 4.85-4.87 (1H, m), 6.10 (1H, br), 6.66 (2H, d, J=9.0Hz), 7.22-7.25 (1H, m), 7.27-7.29 (1H, m), 7.39-7.43 (2H, m), 7.77-7.81(2H, m), 7.85 (1H, d, J=9.0 Hz), 7.96 (1H, d, J=8.7 Hz), 8.03 (2H, d,J=8.7 Hz), 8.18 (1H, d, J=8.7 Hz)

IR (Nujol) 3377, 1621, 1610 cm⁻¹

APCI-MS m/z 563 [M+H]⁺

Synthesis Method of THK-5120

Synthesis of 154

To a tetrahydrofuran (25 ml) suspension of 12 (340 mg, 1.16 mmol), 63(500 mg, 1.39 mmol) and triphenylphosphine (370 mg, 1.39 mmol), atetrahydrofuran (5 ml) solution of diisopropyl azodicarboxylate (0.28ml, 1.39 mmol) was added dropwise over 20 minutes under ice cooling andstirring, and the mixture was stirred at room temperature for 16 hours.To the reaction solution, 63 (500 mg, 1.39 mmol), triphenylphosphine(370 mg, 1.39 mmol) was added and diisopropyl azodicarboxylate (0.28 ml,1.39 mmol) was added under ice cooling and stirring, followed by furtherstirring at room temperature for 3 days. To the reaction solution, 63(170 mg, 0.46 mmol), triphenylphosphine (120 mg, 0.46 mmol) was addedand diisopropyl azodicarboxylate (0.09 ml, 0.46 mmol) was added underice cooling and stirring, followed by further stirring at roomtemperature for 1 day. The solvent of the reaction solution wasdistilled off under reduced pressure and the residue was purified bysilica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/9). To the chloroform (12 ml) solution of theobtained main fraction, trifluoroacetic acid (8 ml) was added dropwiseunder ice cooling and stirring and water (2 ml) was added, followed byfurther stirring at room temperature for 16 hours. To the reactionsolution, ice water was added and the solution was extracted with ethylacetate after adjusting the pH to 8 using an aqueous potassium carbonatesolution. The extraction liquid was dried and the solvent was distilledoff under reduced pressure. The residue was purified by silica gelcolumn chromatography (eluting solvent: ethyl acetate/n-hexane=1/3, 1/1)and then washed with ethyl acetate/n-hexane (1/2) to obtain 154 (410 mg,68%) as pale yellow crystals.

mp 167-168° C.

APCI-MS m/z 521[M+H]⁺

Synthesis of THK-5120

To a methylene chloride (10 ml) solution of 154 (400 mg, 0.77 mmol),3,4-dihydro-2H-pyran (1.39 ml, 15.4 mmol) and paratoluenesulfonic acidmonohydrate (172 mg, 1.00 mmol) were added, and the mixture was stirredat room temperature for 10 minutes. After adjusting the pH of thereaction solution to 9 using triethylamine, the solvent was distilledoff under reduced pressure. The residue was purified by silica gelcolumn chromatography (eluting solvent: ethyl acetate/n-hexane=1/4) toobtain THK-5120 (466 mg, 100%) as a pale yellow amorphous.

¹H NMR (500 MHz, DMSO-d₆) δ 1.14 (6H, t, J=6.9 Hz), 1.34-1.69 (6H, m),2.33, 2.34 (3H, s), 3.22-3.32 (1H, m), 3.42 (4H, q, J=7.3 Hz),3.66-3.72, 3.82-3.87 (1H, m), 4.09-4.37 (5H, m), 4.69-4.72, 4.85-4.87(1H, m), 6.78 (2H, d, J=9.0 Hz), 7.22-7.26 (1H, m), 7.28-7.31 (1H, m),7.38-7.43 (2H, m), 7.77-7.82 (2H, m), 7.86 (1H, d, J=9.0 Hz), 7.97 (1H,d, J=8.7 Hz), 8.07 (2H, d, J=9.0 Hz), 8.20 (1H, d, J=8.3 Hz)

APCI-MS m/z 605[M+H]⁺

Synthesis Method of THK-5121

Synthesis of THK-5121

To a ethanol (12 ml)-acetic acid (1.2 ml)-chloroform (10 ml) solution ofTHK-5119 (600 mg, 1.07 mmol) and an aqueous 20% formaldehyde solution(1.60 ml, 10.7 mmol), picoline borane complex (342 mg, 3.21 mmol) wasadded little by little under ice cooling and stirring, and the mixturewas stirred at room temperature for 1 hour. The reaction solution wasdiluted with ethyl acetate and extracted with ethyl acetate afteradjusting the pH to 8 using an aqueous potassium carbonate solution. Theextraction liquid was washed with water and dried, and then the solventwas distilled off under reduced pressure. The residue was purified bysilica gel column chromatography (eluting solvent: n-hexane/ethylacetate=4/1) and then recrystallized from n-hexane/ethyl acetate toobtain THK-5121 (566 mg, 92%) as pale yellow crystals.

mp 152-154° C.

¹H NMR (500 MHz, DMSO-d₆) δ 1.34-1.68 (6H, m), 2.34 (3H, s), 3.01 (6H,s), 3.41-3.47 (1H, m), 3.65-3.72, 3.81-3.87 (1H, m), 4.10-4.37 (5H, m),4.68-4.72, 4.85-4.87 (1H, m), 6.85 (2H, d, J=7.4 Hz), 7.25-7.37 (2H, m),7.39-7.43 (2H, m), 7.77-7.81 (2H, m), 7.88-7.96 (1H, m), 8.01-8.07 (1H,m), 8.11 (2H, d, J=8.7 Hz), 8.23-8.32 (1H, m)

IR (Nujol) 1622 cm⁻¹

APCI-MS m/z 577[M+H]⁺

Synthesis Method of THK-5122

Synthesis of 156

To a mixture of 115 (650 mg, 1.76 mmol), 6 (580 mg, 2.11 mmol) and1,2-dimethoxyethane (20 ml), sodium carbonate (372 mg, 3.50 mmol) andwater (2 ml) were added and tetrakistriphenylphosphine palladium (102mg, 0.088 mmol) was added under an argon atmosphere and the mixture washeated at reflux for 48 hours. To the reaction solution,tetrakistriphenylphosphine palladium (102 mg, 0.088 mmol) was added, andthe mixture was heated at reflux for 24 hours under an argon atmosphere.The reaction solution was allowed to return to room temperature andsilica gel was added, and then the solvent was distilled off underreduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: n-hexane/ethyl acetate=10/1) to obtain156 (750 mg, 89%) as a pale yellow solid.

mp 104-105° C.

Synthesis of THK-5122

To a mixture of 156 (740 mg, 1.55 mmol) and tetrahydrofuran (20 ml), 1Mtetra-n-butylammonium fluoride/tetrahydrofuran (2.0 ml, 2.00 mmol) wasadded, and the mixture was stirred at room temperature for 1 hour. Thereaction solution was poured into water, and the solution was extractedwith ethyl acetate. The extraction liquid was washed with saturatedsaline and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: n-hexane/ethyl acetate=3/1, 2/1) and thenrecrystallized from ethyl acetate/n-hexane (1/1) to obtain THK-5122 (448mg, 78%) as crystals.

mp 126-127° C.

¹H NMR (400 MHz, CDCl₃) δ 1.21 (6H, t, J=7.0 Hz), 2.60 (1H, brs), 3.43(4H, q, J=7.0 Hz), 4.14-4.23 (2H, m), 4.26-4.37 (1H, m), 4.53-4.62 (1H,m), 4.66-4.74 (1H, m), 6.78 (2H, d, J=8.8 Hz), 7.06 (1H, d, J=2.7 Hz),7.32 (1H, dd, J=9.0, 2.7 Hz), 7.78 (1H, d, J=9.0 Hz), 7.99 (1H, d, J=7.0Hz), 8.01 (1H, d, J=7.0 Hz), 8.03 (2H, d, J=8.8 Hz)

IR (Nujol) 3378, 1620, 1596 cm⁻¹

APCI-MS m/z 369[M+H]⁺

Synthesis Method of THK-5123

Synthesis of 157

To a chloroform (12 ml) solution of 148 (1.20 g, 2.94 mmol),trifluoroacetic acid (8 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 1.5 hours.To the reaction solution, ice water was added, and the solution wasextracted with chloroform/methanol after adjusting the pH to 9 using anaqueous potassium carbonate solution. The extraction liquid was driedthe solvent was distilled off under reduced pressure, and the residuewas purified by silica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/1, ethyl acetate). To a methylene chloride (20 ml)suspension of the obtained orange solid (650 mg) and triethylamine (1.13ml, 8.13 mmol), trifluoroacetic anhydride (0.92 ml, 6.5 mmol) was addeddropwise under ice cooling and stirring, followed by stirring at thesame temperature for 30 minutes. To the reaction solution, ice water wasadded and the solution was extracted with chloroform/methanol. Theextraction liquid was dried the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: ethyl acetate/n-hexane=1/1) and then washed with ethylacetate/n-hexane (1/4) to obtain 157 (360 mg, 37%) as pale browncrystals.

mp 253-254° C.

APCI-MS m/z 333[M+H]⁺

Synthesis of 158

To a tetrahydrofuran (25 ml) suspension of 157 (350 mg, 1.05 mmol), 63(911 mg, 2.53 mmol) and triphenylphosphine (664 mg, 2.53 mmol), atetrahydrofuran (8 ml) solution of diisopropyl azodicarboxylate (0.5 ml,2.52 mmol) was added dropwise over 20 minutes under ice cooling andstirring, and the mixture was stirred at the same temperature for 1hour, followed by stirring at room temperature for 3 days. The solventof the reaction solution was distilled off under reduced pressure, andthe residue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/4). To a chloroform (16 ml) solutionof the obtained main fraction, trifluoroacetic acid (11 ml) was addeddropwise under ice cooling and stirring and water (2.5 ml) was added,followed by further stirring at room temperature for 16 hours. Thereaction solution was diluted with chloroform and extracted withchloroform after adjusting the pH to 8 using an aqueous potassiumcarbonate solution. The extraction liquid was dried and purified bysilica gel column chromatography (eluting solvent: chloroform, ethylacetate/n-hexane=1/2, 1/1) to obtain 158 (490 mg, 83%) as a pale yellowsolid.

mp 186-188° C.

APCI-MS m/z 561[M+H]⁺

Synthesis of 159

To a methylene chloride (20 ml) suspension of 158 (480 mg, 0.86 mmol)and 3,4-dihydro-2H-pyran (1.55 ml, 17.2 mmol), paratoluenesulfonic acidmonohydrate (192 mg, 1.12 mmol) was added, and the mixture was stirredat room temperature for 10 minutes. After adjusting the pH of thereaction solution to 8 by adding triethylamine, the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/4,1/2) to obtain 159 (480 mg, 87%) as a pale yellow solid.

mp 112-115° C.

APCI-MS m/z 645[M+H]⁺

Synthesis of THK-5123

To a tetrahydrofuran (9 ml)-water (3 ml) solution of 159 (470 mg, 0.73mmol), lithium hydroxide monohydrate (46 mg, 1.09 mmol) was added underice cooling and stirring, and the mixture was stirred at the sametemperature for 1.5 hours, followed by stirring at room temperature for1 hour. Lithium hydroxide monohydrate (46 mg, 1.09 mmol) and methanol (2ml) were added, followed by further stirring at room temperature for 16hours. The reaction solution was diluted with ethyl acetate, washed withwater and dried, and the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: ethyl acetate/n-hexane=2/3, 1/1) to obtain THK-5123(380 mg, 95%) as a pale yellow amorphous.

¹H NMR (500 MHz, DMSO-d₆) δ 1.34-1.71 (6H, m), 2.33, 2.34 (3H, s),3.37-3.46 (1H, m). 3.66-3.72, 3.82-3.87 (1H, m), 4.08-4.36 (5H, m),4.69-4.72, 4.84-4.87 (1H, m), 5.52 (2H, br), 6.68 (2H, d, J=8.7 Hz),7.21-7.25 (1H, m), 7.26-7.29 (1H, m), 7.38-7.43 (2H, m), 7.77-7.81 (2H,m), 7.84 (1H, d, J=9.3 Hz), 7.93 (1H, d, J=8.7 Hz), 7.96 (1H, d, J=8.7Hz), 8.17 (1H, d, J=8.7 Hz)

IR (Nujol) 3452, 1728, 1622 cm⁻¹

APCI-MS m/z 549[M+H]⁺

Synthesis Method of THK-5125

Synthesis of 161

To a mixture of 115 (650 mg, 1.76 mmol), 160 (733 mg, 2.11 mmol) and1,2-dimethoxyethane (20 ml), sodium carbonate (372 mg, 3.50 mmol) andwater (2 ml) were added, and tetrakistriphenylphosphine palladium (102mg, 0.088 mmol) was added under an argon atmosphere, and then themixture was heated at reflux for 33 hours. The reaction solution wasallowed to return to room temperature, filtered with celite and thenpurified by silica gel flash column chromatography (eluting solvent:n-hexane/ethyl acetate=10/1) to obtain 161 (900 mg, 93%) as a viscousoily substance.

Synthesis of THK-5125

To a mixture of 161 (900 mg, 1.62 mmol) and tetrahydrofuran (20 ml), 1Mtetra-n-butylammonium fluoride/tetrahydrofuran (2.0 ml, 2.00 mmol) wasadded, and the mixture was stirred at room temperature for 1 hour. Thereaction solution was poured into water, and the solution was extractedwith ethyl acetate. The extraction liquid was washed with saturatedsaline and dried, and then the solvent was distilled off under reducedpressure. To a methylene chloride (20 ml) solution of the residue,trifluoroacetic acid (4 ml) was added dropwise, followed by stirring atroom temperature for 1.5 hours. The reaction solution was diluted withethyl acetate and the solution was made basic with an aqueous saturatedsodium hydrogen carbonate solution. The organic layer was washed withsaturated saline and dried, and then the solvent was distilled off underreduced pressure. The residue was recrystallized from ethyl acetate toobtain THK-5125 (350 mg, 63%) as pale brown crystals.

mp 146-147° C.

¹H NMR (400 MHz, CDCl₃) δ 1.29 (3H, t, J=7.1 Hz), 2.61 (1H, br), 3.24(2H, q, J=7.1 Hz), 3.80 (1H, br), 4.14-4.23 (2H, m), 4.32 (1H, brd, J=18Hz), 4.54-4.62 (1H, m), 4.65-4.74 (1H, m), 6.71 (2H, d, J=8.8 Hz), 7.07(1H, d, J=2.7 Hz), 7.33 (1H, dd, J=9.0, 2.7 Hz), 7.76 (1H, d, J=8.8 Hz),8.01 (2H, d, J=8.8 Hz), 8.02 (1H, d, J=9.0 Hz), 7.90-8.20 (1H, br)

IR (Nujol) 3432, 3356, 1621, 1598 cm⁻¹

APCI-MS m/z 341 [M+H]⁺

Synthesis Method of THK-5131

Synthesis of 162

To a 1,2-dimethoxyethane (20 ml) solution of 147 (600 mg, 2.38 mmol) and160 (830 mg, 2.38 mmol), potassium carbonate (990 mg, 7.14 mmol), water(0.41 ml) and tetrakistriphenylphosphine palladium (275 mg, 0.24 mmol)were added under an argon atmosphere, and the mixture was stirred at 80°C. for 16 hours. The reaction solution was allowed to return to roomtemperature and water was added, and the solution was extracted withethyl acetate. The extraction liquid was washed with water and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/9) to obtain 162 (1.02 g, 98%) as apale yellow viscous oily substance.

APCI-MS m/z 437[M+H]⁺

Synthesis of 163

To a chloroform (10 ml) solution of 162 (1.01 g, 2.31 mmol),trifluoroacetic acid (1 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at the same temperature for 40minutes and trifluoroacetic acid (0.5 ml) was added, followed bystirring at the same temperature for 10 minutes. To the reactionsolution, ice water-ethyl acetate was added, and the solution wasextracted with ethyl acetate after adjusting the pH to 9 using anaqueous potassium carbonate solution. The extraction liquid was driedand the solvent was distilled off under reduced pressure and the residuewas purified by silica gel column chromatography (eluting solvent:n-hexane/ethyl acetate=4/1, 2/1) to obtain 163 (740 mg, 88%) as a paleyellow solid.

mp 205-206° C.

APCI-MS m/z 365[M+H]⁺

Synthesis of 164

To a tetrahydrofuran (30 ml) solution of 163 (730 mg, 2.00 mmol), 63(1.74 g, 4.81 mmol) and triphenylphosphine (1.26 g, 4.81 mmol), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (0.95ml, 4.81 mmol) was added dropwise over 20 minutes under ice cooling andstirring, and the mixture was stirred at the same temperature for 1hour, followed by stirring at room temperature for 4 days. The solventof the reaction solution was distilled off under reduced pressure, andthe residue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/9). To a chloroform (24 ml) solutionof the obtained colorless oily substance (2.53 g), trifluoroacetic acid(16 ml) was added dropwise under ice cooling and stirring and water (4ml) was added, followed by stirring at room temperature for 16 hours. Tothe reaction solution, ice water and ethyl acetate were added and thesolution was extracted with ethyl acetate by adjusting the pH to 8 usingan aqueous potassium carbonate solution. The extraction liquid was driedand the solvent was distilled off under reduced pressure. The residuewas purified by silica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/2, ethyl acetate) and then washed with ethylacetate/n-hexane (1/2) to obtain 164 (880 mg, 89%) as yellow crystals.

mp 162-164° C.

APCI-MS m/z 493[M+H]⁺

Synthesis of THK-5131

To a methylene chloride (20 ml) suspension of 164 (440 mg, 0.893 mmol),3,4-dihydro-2H-pyran (1.62 ml, 17.9 mmol) and paratoluenesulfonic acidmonohydrate (200 mg, 1.16 mmol) were added, and the mixture was stirredat room temperature for 15 minutes. After adjusting the pH of thereaction solution to 8 using triethylamine, the solvent was distilledoff under reduced pressure. The residue was purified by silica gelcolumn chromatography (eluting solvent: ethyl acetate/n-hexane=1/2) andthen recrystallized from ethyl acetate/n-hexane to obtain THK-5131 (389mg, 76%) as pale yellow crystals.

mp 103-105° C.

¹H NMR (500 MHz, DMSO-d₆) δ 1.20 (3H, t, J=7.1 Hz), 1.34-1.69 (6H, m),2.33, 2.34 (3H, s), 3.09-3.15 (2H, m), 3.37-3.46 (1H, m), 3.66-3.72,3.80-3.88 (1H, m), 4.09-4.37 (5H, m), 4.69-4.72, 4.84-4.87 (1H, m), 6.01(1H, br), 6.67 (2H, d, J=8.3 Hz), 7.23 (1H, d, J=9.0 Hz), 7.26-7.29 (1H,m), 7.38-7.43 (2H, m), 7.77-7.81 (2H, m), 7.85 (1H, d, J=9.0 Hz), 7.95(1H, d, J=8.7 Hz), 8.01 (2H, d, J=8.7 Hz), 8.18 (1H, d, J=8.0 Hz)

IR (Nujol) 3360, 1619, 1608 cm⁻¹

APCI-MS m/z 577[M+H]⁺

Synthesis Method of THK-5127

Synthesis of 165

To a 1,2-dimethoxyethane (63 ml) suspension of 147 (1.94 g, 7.71 mmol)and 94 (2.72 g, 8.48 mmol), potassium carbonate (3.20 g, 23.1 mmol),water (1.34 ml) and tetrakistriphenylphosphine palladium (890 mg, 0.77mmol) were added under an argon atmosphere, and the mixture was stirredat 80° C. for 16 hours. The reaction solution was allowed to return toroom temperature and water was added, and the solution was extractedwith ethyl acetate. The extraction liquid was washed with water anddried, and then the solvent was distilled off under reduced pressure.The residue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/4) and then recrystallized from ethylacetate/n-hexane to obtain 165 (2.78 g, 88%) as pale yellow crystals.

mp 182-183° C.

APCI-MS m/z 410[M+H]⁺

Synthesis of 166

To a chloroform (40 ml) solution of 165 (2.77 g, 6.76 mmol),trifluoroacetic acid (4 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 20minutes. To the reaction solution, ice water was added and the solutionwas extracted with ethyl acetate-tetrahydrofuran after adjusting the pHto 9 using an aqueous potassium carbonate solution. The extractionliquid was dried and the solvent was distilled off under reducedpressure, and then the residue was recrystallized from ethyl acetate toobtain 166 (2.00 g, 88%) as pale yellow crystals.

mp 325-326° C.

APCI-MS m/z 338[M+H]⁺

Synthesis of 167

To a tetrahydrofuran (40 ml) solution of 166 (1.50 g, 4.45 mmol), 16(1.11 g, 5.35 mmol) and triphenylphosphine (1.40 g, 5.35 mmol), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (1.06ml, 5.35 mmol) was added dropwise over 30 minutes under ice cooling andstirring, and the mixture was stirred at the same temperature for 1hour, followed by stirring at room temperature for 3 days. To thereaction solution, 16 (830 mg, 4.00 mmol) and triphenylphosphine (1.05g, 4.00 mmol) were additionally added, and the diisopropylazodicarboxylate (0.79 ml, 4.00 mmol) was added dropwise under icecooling and stirring, followed by further stirring at room temperaturefor 5 hours. The solvent of the reaction solution was distilled offunder reduced pressure, and the residue was purified by silica gelcolumn chromatography (eluting solvent: ethyl acetate/n-hexane=1/9, 1/4)and then recrystallized from ethyl acetate/n-hexane to obtain 167 (1.51g, 64%) as colorless crystals.

mp 188-189° C.

APCI-MS m/z 528[M+H]⁺

Synthesis of THK-5127

To a chloroform (24 ml) solution of 167 (1.50 g, 2.84 mmol),trifluoroacetic acid (16 ml) was added dropwise under ice cooling andstirring, and water (4 ml) was added, and the mixture was stirred atroom temperature for 16 hours. To the reaction solution, ice water andthen ethyl acetate were added, and the solution was extracted with ethylacetate after adjusting the pH to 9 using an aqueous potassium carbonatesolution. The extraction liquid was dried and the solvent was distilledoff under reduced pressure. The residue was purified by silica gelcolumn chromatography (eluting solvent: ethyl acetate) and thenrecrystallized from ethyl acetate to obtain THK-5127 (717 mg, 81%) aspale yellow crystals.

mp 196-197° C.

¹H NMR (400 MHz, DMSO-d₆) δ 4.06-4.19 (3H, m), 4.44-4.65 (2H, m), 5.54(1H, d, J=5.7 Hz), 6.33 (2H, s), 6.57 (1H, d, J=8.8 Hz), 7.36-7.41 (2H,m), 7.87-7.91 (1H, m), 7.97 (1H, d, J=8.8 Hz), 8.21-8.27 (2H, m), 8.80(1H, d, J=2.4 Hz)

IR (Nujol) 3440, 1651, 1619 cm⁻¹

APCI-MS m/z 314[M+H]⁺

Synthesis Method of THK-5150

Synthesis of 168

To a tetrahydrofuran (30 ml) solution, 166 (980 mg, 2.91 mmol), 63 (2.51g, 6.98 mmol) and triphenylphosphine (1.83 g, 6.98 mmol), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (1.38ml, 6.98 mmol) was added dropwise over 10 minutes under ice cooling andstirring, and the mixture was stirred at the same temperature for 1hour, followed by stirring at room temperature for 2 days. The solventof the reaction solution was distilled off under reduced pressure, andthe residue was washed with ethyl acetate/n-hexane=1/4) after grindingto obtain 168 (1.38 g, 70%) as colorless crystals.

mp 204-206° C.

APCI-MS m/z 680 [M+H]⁺

Synthesis of 169

To a chloroform (24 ml) solution of 168 (1.98 g, 2.91 mmol),trifluoroacetic acid (16 ml) was added dropwise under ice cooling andstirring, and water (4 ml) was added, and the mixture was stirred atroom temperature for 16 hours. To reaction solution, ice water and ethylacetate were added, and the solution was extracted with ethyl acetateafter adjusting the pH to 8 using an aqueous potassium carbonatesolution, and the solution was extracted with ethyl acetate afteradjusting the pH to 8 using an aqueous potassium carbonate solution. Theextraction liquid was dried and the solvent was distilled off underreduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/1, ethylacetate) and then washed with ethyl acetate/n-hexane (1/1) to obtain 169(940 mg, 69%) as colorless crystals.

mp 168-169° C.

APCI-MS m/z 466[M+H]⁺

Synthesis of THK-5150

To a ethylene chloride (20 ml) solution of 169 (930 mg, 2.00 mmol),3,4-dihydro-2H-pyran (3.62 ml, 40.0 mmol) and paratoluenesulfonic acidmonohydrate (447 mg, 2.60 mmol) were added, and the mixture was stirredat room temperature for 40 minutes. Paratoluenesulfonic acid monohydrate(344 mg, 2.0 mmol) was added, followed by stirring at room temperaturefor 10 minutes. After adjusting the pH of the reaction solution to 8 byadding triethylamine, ethyl acetate and water were added, followed bystirring at room temperature for 10 minutes. The organic layer wasseparated and dried, and then the solvent was distilled off underreduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent:ethyl acetate/n-hexane=1/1, 2/1, ethylacetate) to obtain THK-5150 (510 mg, 46%) as a colorless foam-likesubstance.

¹H NMR (500 MHz, DMSO-d₆) δ 1.35-1.68 (6H, m), 2.34, 2.34 (3H, s),3.36-3.46 (1H, m). 3.66-3.71, 3.82-3.87 (1H, m), 4.09-4.37 (5H, m),4.70, 4.84-4.87 (1H, m), 6.37 (2H, s), 6.57 (1H, d, J=8.7 Hz), 7.23-7.27(1H, m), 7.29-7.31 (1H, m), 7.41 (2H, dd, J=8.2, 1.8 Hz), 7.79 (2H, dd,J=8.7, 2.6 Hz), 7.87 (1H, d, J=9.0 Hz), 7.98 (1H, d, J=8.7 Hz), 8.21(1H, d, J=8.7 Hz), 8.25 (1H, dd, J=8.7, 2.6 Hz), 8.80 (1H, d, J=2.2 Hz)

IR (Nujol) 1621 cm⁻¹

APCI-MS m/z 550[M+H]⁺

Synthesis Method of THK-5152

Synthesis of 170

To an N,N-dimethylformamide (30 ml) suspension of 165 (1.87 g, 4.57mmol), 60% sodium hydride (220 mg, 5.5 mmol) was added little by littleunder ice cooling and stirring an argon atmosphere, and the mixture wasstirred at the same temperature for 15 minutes and methyl iodide (0.43ml, 6.9 mmol) was added dropwise at the same temperature, followed bystirring at the same temperature for 5 minutes and further stirring atroom temperature for 30 minutes. To the reaction solution, ice water wasadded and the solution was extracted with ethyl acetate. The extractionliquid was washed with water and dried, and then the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/9,1/4) to obtain 170 (1.93 g, 100%) as a pale yellow oily substance.

APCI-MS m/z 424[M+H]⁺

Synthesis of 171

To chloroform (20 ml) solution of 170 (1.93 g, 4.56 mmol),trifluoroacetic acid (4 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at the same temperature for 1hour. To the reaction solution, water and ethyl acetate were added, andthe solution was extracted with ethyl acetate after adjusting the pH to8 using an aqueous potassium carbonate solution. The extraction liquidwas dried and the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: n-hexane/ethyl acetate=2/1) and then washed with n-hexane/ethylacetate=2/1 to obtain 171 (1.57 g, 98%) as pale yellow crystals.

mp 179-180° C.

APCI-MS m/z 352[M+H]⁺

Synthesis of 172

To a tetrahydrofuran (20 ml) suspension of 171 (500 mg, 1.42 mmol), 63(1.23 g, 3.42 mmol) and triphenylphosphine (900 mg, 3.42 mmol), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (0.68ml, 3.42 mmol) was added dropwise over 5 minutes under ice cooling andstirring, and the mixture was stirred at the same temperature for 1hour, followed by stirring at room temperature for 16 hours. The solventof the reaction solution was distilled off under reduced pressure, andthe residue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/9, 1/4) and then recrystallized fromethyl acetate/n-hexane (1/9) to obtain 172 (950 mg, 96%) as colorlesscrystals.

mp 112-113° C.

APCI-MS m/z 694[M+H]⁺

Synthesis of 173

To a chloroform (18 ml) solution of 172 (940 mg, 1.35 mmol),trifluoroacetic acid (12 ml) was added dropwise under ice cooling andstirring, and water (3 ml) was added, and the mixture was stirred atroom temperature for 16 hours. To the reaction solution, ice water andethyl acetate were added, and the solution was extracted with ethylacetate after adjusting the pH to 8 using an aqueous potassium carbonatesolution. The extraction liquid was dried and the solvent was distilledoff under reduced pressure, and the residue was purified by silica gelcolumn chromatography (eluting solvent: ethyl acetate/n-hexane=1/1,ethyl acetate) to obtain 173 (400 mg, 62%) as a pale yellow solid.

mp 162-163° C.

APCI-MS m/z 480[M+H]⁺

Synthesis of THK-5152

To a chloroform (20 ml) suspension of 173 (390 mg, 0.81 mmol),3,4-dihydro-2H-pyran (1.47 ml, 16.3 mmol) and paratoluenesulfonic acidmonohydrate (322 mg, 1.87 mmol) were added, and the mixture was stirredat room temperature for 20 minutes. After adjusting the pH of thereaction solution to 8 by adding triethylamine, the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=2/1,ethyl acetate) and then recrystallized from ethyl acetate/n-hexane toobtain THK-5152 (442 mg, 97%) as colorless crystals.

mp 129-130° C.

¹H NMR (400 MHz, DMSO-d₆) δ 1.33-1.71 (6H, m), 2.32 (3H, s), 2.85 (3H,d, J=4.8 Hz), 3.36-3.51 (1H, m), 3.65-3.73, 3.81-3.89 (1H, m), 4.09-4.38(5H, m), 4.70, 4.86 (1H, m), 6.58 (1H, d, J=8.8 Hz), 6.89 (1H, q, J=4.8Hz), 7.23-7.27 (1H, m), 7.29-7.32 (1H, m), 7.39-7.43 (2H, m), 7.79 (2H,dd, J=8.3, 2.0 Hz), 7.87 (1H, d, J=9.1 Hz), 7.99 (1H, d, J=8.8 Hz), 8.21(1H, d, J=8.8 Hz), 8.26 (1H, dd, J=8.8, 2.4 Hz), 8.87 (1H, d, J=2.4 Hz)

IR (Nujol) 3355, 1623, 1604 cm⁻¹

APCI-MS m/z 564[M+H]⁺

Synthesis Method of THK-5129

Synthesis of THK-5129

To a methanol (8.6 ml)-acetic acid (0.86 ml) solution THK-of 5127 (123mg, 0.39 mmol) and an aqueous 20% formaldehyde solution (1.18 ml, 7.8mmol), a picoline borane complex (252 mg, 2.36 mmol) was added little bylittle under ice cooling and stirring, and the mixture was stirred atroom temperature for 3 hours. To the reaction solution, water was added,and the solution was extracted with ethyl acetate. The extraction liquidwas washed with water and dried, and then the solvent was distilled offunder reduced pressure. The residue was purified by NH silica gel columnchromatography (eluting solvent: n-hexane/ethyl acetate=1/2), purifiedby silica gel column chromatography (eluting solvent: n-hexane/ethylacetate=1/2) and then recrystallized from ethyl acetate/n-hexane toobtain THK-5129 (100 mg, 75%) as pale yellow crystals.

mp 172-173° C.

¹H NMR (400 MHz, DMSO-d₆) δ 3.12 (6H, s), 4.06-4.19 (3H, m), 4.44-4.65(2H, m), 5.54 (1H, d, J=4.8 Hz), 6.78 (1H, d, J=9.1 Hz), 7.37-7.41 (2H,m), 7.88-7.93 (1H, m), 8.01 (1H, d, J=8.8 Hz), 8.25 (1H, d, J=8.8 Hz),8.36 (1H, dd, J=9.1, 2.4 Hz), 8.95 (1H, d, J=2.4 Hz)

IR (Nujol) 3378, 1620, 1611 cm⁻¹

APCI-MS m/z 342[M+H]⁺

Synthesis Method of THK-5135

Synthesis of THK-5135

To a methanol (20 ml)-acetic acid (2 ml) solution of THK-5150 (500 mg,0.91 mmol) and an aqueous 20% formaldehyde solution (2.73 ml, 18.2mmol), a picoline borane complex (584 mg, 5.46 mmol) was added little bylittle at room temperature under stirring, and the mixture was stirredat room temperature for 1 hour. To the reaction solution, water andethyl acetate were added, and the solution was extracted with ethylacetate after adjusting the pH to 9 using an aqueous potassium carbonatesolution. The extraction liquid was washed with water and dried, andthen the solvent was distilled off under reduced pressure. The residuewas purified by silica gel column chromatography (eluting solvent:n-hexane/ethyl acetate=1/1) and then recrystallized from ethylacetate/n-hexane to obtain THK-5135 (331 mg, 63%) as pale yellowcrystals.

mp 135-138° C.

¹H NMR (400 MHz, DMSO-d₆) δ 1.34-1.70 (6H, m), 2.34 (3H, s), 3.12 (6H,s), 3.38-3.51 (1H, m), 3.65-3.73, 3.81-3.89 (1H, m), 4.08-4.38 (5H, m),4.69-4.73, 4.84-4.88 (1H, m), 6.79 (1H, d, J=9.1 Hz), 7.24-7.28 (1H, m),7.30-7.33 (1H, m), 7.39-7.43 (2H, m), 7.79 (2H, dd, J=8.5, 2.1 Hz), 7.88(1H, d, J=9.1 Hz), 8.02 (1H, d, J=8.8 Hz), 8.23 (1H, d, J=8.8 Hz), 8.37(1H, dd, J=9.1, 2.4 Hz), 8.96 (1H, d, J=2.4 Hz)

IR (Nujol) 1624 cm⁻¹

APCI-MS m/z 578[M+H]⁺

Synthesis Method of THK-5130

Synthesis of THK-5130

To a methanol (16 ml)-acetic acid (1.6 ml) suspension of THK-5127 (449mg, 1.43 mmol) and acetaldehyde (78 mg/ml methanol solution: 3.24 ml,5.73 mmol), a picoline borane complex (460 mg, 4.30 mmol) was addedlittle by little under ice cooling and stirring, and the mixture wasstirred at room temperature for 4 hours. Acetaldehyde (78 mg/ml methanolsolution: 3.24 ml, 5.73 mmol) and a picoline borane complex (460 mg,4.30 mmol) were added, followed by stirring at room temperature for 16hours. To the reaction solution, water and ethyl acetate were added, andthe solution was extracted with ethyl acetate after adjusting the pH to9 using an aqueous potassium carbonate solution. The extraction liquidwas washed with water and dried, and then the solvent was distilled offunder reduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: n-hexane/ethyl acetate=2/1, 1/1) andthen recrystallized from ethyl acetate/n-hexane to obtain THK-5130 (479mg, 91%) as pale yellow crystals.

mp 131-132° C.

¹H NMR (400 MHz, DMSO-d₆) δ 1.15 (6H, t, J=7.1 Hz), 3.57 (4H, q, J=7.1Hz), 4.00-4.19 (3H, m), 4.44-4.65 (2H, m), 5.54 (1H, d, J=4.8 Hz), 6.74(1H, d, J=9.1 Hz), 7.36-7.41 (2H, m), 7.87-7.92 (1H, m), 8.00 (1H, d,J=8.8 Hz), 8.24 (1H, d, J=8.8 Hz), 8.33 (1H, dd, J=9.1, 2.4 Hz), 8.93(1H, d, J=2.4 Hz)

IR (Nujol) 3275, 1622 cm⁻¹

APCI-MS m/z 370[M+H]⁺

Synthesis Method of THK-5138

Synthesis of 174

To a methanol (13 ml)-acetic acid (1.3 ml)-chloroform (10 ml) solutionof 169 (550 mg, 1.18 mmol) and acetaldehyde (0.66 ml, 11.8 mmol),picoline borane complex (890 mg, 8.32 mmol) was added little by littleunder ice cooling and stirring, and the mixture was stirred at roomtemperature for 4 days. To the reaction solution, water and ethylacetate were added, and the solution was extracted with ethyl acetateafter adjusting the pH to 8 using an aqueous potassium carbonatesolution. The extraction liquid was washed with water and dried, andthen the solvent was distilled off under reduced pressure. The residuewas purified by silica gel column chromatography (eluting solvent:n-hexane/ethyl acetate=1/1, ethyl acetate) to obtain 174 (560 mg, 91%)as a pale yellow foam-like substance.

APCI-MS m/z 522[M+H]⁺

Synthesis of THK-5138

To a chloroform (20 ml) solution of 174 (550 mg, 1.05 mmol),3,4-dihydro-2H-pyran (1.19 ml, 21.1 mmol), paratoluenesulfonic acidmonohydrate (420 mg, 2.43 mmol) was added, and the mixture was stirredat room temperature for 10 minutes. After adjusting the pH of thereaction solution to 8 by adding triethylamine, the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/4,1/2) and then washed with ethyl acetate/n-hexane to obtain THK-5138 (477mg, 75%) as pale yellow crystals.

mp 102-104° C.

¹H NMR (400 MHz, DMSO-d₆) δ 1.15 (6H, t, J=7.0 Hz), 1.30-1.50 (4H, m),1.52-1.69 (2H, m), 2.34 (3H, s), 3.36-3.46 (1H, m), 3.57 (4H, q, J=6.7Hz), 3.65-3.73, 3.81-3.88 (1H, m), 4.08-4.38 (5H, m), 4.69-4.72,4.84-4.88 (1H, m), 6.73 (1H, d, J=9.1 Hz), 7.23-7.27 (1H, m), 7.29-7.32(1H, m), 7.41 (2H, d, J=7.3 Hz), 7.79 (2H, dd, J=8.2, 2.1 Hz), 7.87 (1H,d, J=9.1 Hz), 8.00 (1H, d, J=8.8 Hz), 8.22 (1H, d, J=8.8 Hz), 8.34 (1H,dd, J=9.1, 2.4 Hz), 8.94 (1H, d, J=2.4 Hz)

IR (Nujol) 1620 cm⁻¹

APCI-MS m/z 606[M+H]⁺

Synthesis Method of THK-5151

Synthesis of 175

To 115 (175 mg, 0.473 mmol), 99 (71.1 mg, 0.505 mmol), sodium carbonate(105 mg, 0.992 mmol) and dichlorobis(triphenylphosphine)palladium (3.6mg, 0.00513 mmol), 2.5 ml of a mixed solution ofwater/ethanol/1,2-dimethoxyethane (each ratio=0.55:0.4:1.25) was added,and the mixture was heated at reflux at 90° C. for 90 minutes. Thereaction solution was allowed to return to room temperature and water(40 ml) was added, and the solution was extracted with ethyl acetate(2×40 ml) and then dried over magnesium sulfate. The extraction solventwas distilled off and the residue was purified by silica gel flashcolumn chromatography (eluting solvent: n-hexane/ethyl acetate=85/15,60/40) to obtain 175 (191 mg, 94%) as crystalline substance. E1-MS m/z430 [M]+

Synthesis of 176

175 (106 mg, 0.247 mmol) was dissolved in tetrahydrofuran (3.0 ml) and a1M tetrabutylammonium fluoride/acetonitrile solution (0.37 ml, 0.37mmol) was added, and the mixture was reacted at room temperature for 90minutes reaction. To the reaction solution, water (30 ml) was added, andthe solution was extracted with ethyl acetate (3×30 ml) and then driedover magnesium sulfate. The extraction solvent was distilled off, andthe residue was purified by silica gel flash column chromatography(eluting solvent: n-hexane/ethyl acetate=45/55, 20/80) to obtain 176(74.1 mg, 95%) as white crystals. EI-MS m/z 316[M]⁺.

Synthesis of THK-5151

176 (52.4 mg, 0.166 mmol) was dissolved in methanol (2.0 ml) and a 40%monomethylaamine/methanol solution (0.60 ml, 5.8 mmol) was added, andthe mixture was heated at 40° C. for 7 hours, followed by stirring withheating at 50° C. for 17 hours. After the reaction solution returned toroom temperature, the solvent was distilled off and the residue waspurified by silica gel flash column chromatography (eluting solvent:ethyl acetate/methanol=100/0, 70/30) to obtain THK-5151 (46.0 mg, 85%)as yellow crystals. EI-MS m/z 327 [M]⁺.

¹H NMR (600 MHz, CDCl₃) δ 3.01 (3H, d, J=4.8 Hz), 4.20-4.25 (2H, m),4.32-4.38 (1H, m), 4.59-4.65 (1H, m), 4.67-4.73 (1H, m), 4.85 (1H, br),6.54 (1H, d, J=8.4 Hz), 7.11 (1H, d, J=3.0 Hz), 7.37 (1H, dd, J=3.0, 9.6Hz), 7.76 (1H, d, J=8.4 Hz), 8.02 (1H, d, J=9.6 Hz), 8.06 (1H, d, J=8.4Hz), 8.36 (1H, dd, J=1.8, 9.0 Hz), 8.82 (1H, d, J=1.8 Hz).

Synthesis Method of THK-5177

Synthesis of 189

From 115 (112 mg, 0.302 mmol) and 188 (91.0 mg, 0.301 mmol), 189 (ayellow crystal, 94.6 mg, 62%) was synthesized in the same manner as inthe compound 175. EI-MS m/z 509[M]⁺.

Synthesis of THK-5177

189 (75.0 mg, 0.147 mmol) was dissolved in tetrahydrofuran (3.0 ml) anda 1M tetrabutylammonium fluoride/acetonitrile solution (0.37 ml, 0.37mmol) was added, and the mixture was reacted at room temperature for 90minutes reaction. To the reaction solution, water (30 ml) was added, andthe solution was extracted with ethyl acetate (3×30 ml) and then driedover magnesium sulfate. The extraction solvent was distilled off and theresidue was purified by silica gel flash column chromatography (elutingsolvent: n-hexane/ethyl acetate=45/55, 20/80) to synthesize THK-5177(yellow crystals, 40.8 mg, 70%). EI-MS m/z 395[M]⁺.

¹H NMR (600 MHz, CDCl₃) δ 2.38 (3H, s), 2.54 (1H, br), 2.61 (4H, t,J=4.8 Hz), 3.33 (4H, t, J=4.8 Hz), 4.20-4.24 (2H, m), 4.31-4.38 (1H, m),4.59-4.65 (1H, m), 4.67-4.73 (1H, m), 7.04 (1H, d, J=9.0 Hz), 7.10 (1H,d, J=3.0 Hz), 7.36 (1H, dd, J=3.0, 9.0 Hz), 7.81 (1H, d, J=8.4 Hz), 8.04(1H, d, J=9.6 Hz), 8.05 (1H, d, J=9.0 Hz), 8.08 (2H, d, J=9.0 Hz).

Synthesis Method of THK-5178

Synthesis of 191

From 115 (134 mg, 0.361 mmol) and 190 (146 mg, 0.505 mmol), 191 (ayellow crystal, 144 mg, 80%) was synthesized in the same manner as inthe compound

175. EI-MS m/z 496[M]⁺.

Synthesis of THK-5178

From 191 (30.4 mg, 0.065 mmol), THK-5178 (a milk white crystal, 13.3 mg,54%) was synthesized in the same manner as in THK-5177. EI-MS m/z 382[M]⁺. ¹H NMR (600 MHz, CDCl₃) δ 2.19 (1H, s), 2.54 (1H, br), 3.02 (4H,t, J=5.4 Hz), 3.63 (4H, t, J=5.4 Hz), 4.20-4.24 (2H, m), 4.32-4.38 (1H,m), 4.59-4.65 (1H, m), 4.67-4.73 (1H, m), 6.78 (1H, d, J=9.0 Hz), 7.11(1H, d, J=2.4 Hz), 7.37 (1H, dd, J=2.4, 9.0 Hz), 7.78 (1H, d, J=9.0 Hz),8.02 (1H, d, J=9.6 Hz), 8.06 (1H, d, J=8.4 Hz), 8.38 (1H, dd, J=2.4, 9.0Hz), 8.90 (1H, d, J=2.4 Hz).

Synthesis Method of THK-5180

Synthesis of 195

From 115 (180 mg, 0.486 mmol) and 194 (157 mg, 0.517 mmol), 195 (a whitecrystal, 119 mg, 48%) was synthesized in the same manner as in thecompound

175. EI-MS m/z 510[M]⁺.

Synthesis of THK-5180

From 195 (86.0 mg, 0.168 mmol), THK-5180 (white crystals, 48.6 mg, 73%)was synthesized in the same manner as in THK-5177. EI-MS m/z 396[M]⁺.

¹1H NMR (600 MHz, CDCl₃) δ 2.37 (3H, s), 2.46 (1H, d, J=5.4 Hz), 2.55(4H, t, J=5.4 Hz), 3.69 (4H, t, J=5.4 Hz), 4.20-4.25 (2H, m), 4.31-4.38(1H, m), 4.59-4.65 (1H, m), 4.67-4.73 (1H, m), 6.79 (1H, d, J=9.0 Hz),7.11 (1H, d, J=2.4 Hz), 7.37 (1H, dd, J=3.0, 9.6 Hz), 7.76 (1H, d, J=9.0Hz), 8.02 (1H, d, J=9.0 Hz), 8.06 (1H, d, J=8.4 Hz), 8.38 (1H, dd,J=2.4, 9.0 Hz), 8.90 (1H, d, J=2.4 Hz).

Synthesis Method of THK-5142

200

To an N,N-dimethylformamide (30 ml) suspension of 165 (2.09 g, 5.10mmol), 60% sodium hydride (245 mg, 6.12 mmol) was added little by littleunder ice cooling and stirring and an argon atmosphere. After stirringat the same temperature for 10 minutes, ethyl iodide (0.616 ml, 7.66mmol) was added dropwise, and the mixture was stirred at the sametemperature for 1 hour, followed by stirring at room temperature for 30minutes. To the reaction solution, ice water was added and the solutionwas extracted with ethyl acetate. The extraction liquid was washed withwater and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: ethyl acetate/n-hexane=1/4) to obtain 200 (2.07 g,92%) as a colorless oily substance.

APCI-MS m/z 438[M+H]⁺

201

To a chloroform (30 ml) solution of 200 (2.06 g, 4.71 mmol),trifluoroacetic acid (5 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at the same temperature for 20minutes. To the reaction solution, water was added, and the solution wasextracted with ethyl acetate after adjusting the pH to 7 using anaqueous potassium carbonate solution. The extraction liquid was driedand the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (elutingsolvent: n-hexane/ethyl acetate=2/1) and then recrystallized fromn-hexane/ethyl acetate to obtain 201 (1.26 g, 73%) as pale yellowcrystals.

mp 181-182° C.

APCI-MS m/z 366 [M+H]+

Synthesis of THK-5142

To a tetrahydrofuran (20 ml) solution of 201 (750 mg, 2.05 mmol), 16(1.03 g, 4.93 mmol) and triphenylphosphine (1.29 g, 4.93 mmol), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (0.98ml, 4.93 mmol) was added dropwise under ice cooling and stirring, andthe mixture was stirred at the same temperature for 1 hour, followed bystirring at room temperature for 16 hours. The solvent of the reactionsolution was distilled off under reduced pressure, and the residue waspurified by silica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/20, 1/9) to obtain a colorless oily substance (1.69g) containing the objective product. To a chloroform (18 ml) solution ofthe present product (1.69 g), trifluoroacetic acid (12 ml) was addeddropwise under ice cooling and stirring, and water (3 ml) was added,followed by stirring at room temperature for 16 hours. To the reactionsolution, ice water and then ethyl acetate were added, and the solutionwas extracted with ethyl acetate after adjusting the pH to 9 using anaqueous potassium carbonate solution. The extraction liquid was driedand the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/1, 2/1) and then recrystallized fromethyl acetate to obtain THK-5142 (465 mg, 66%) as colorless crystals.

mp 154-155° C.

¹H NMR (400 MHz, DMSO-d₆) δ 1.17 (3H, t, J=7.3 Hz), 3.29-3.38 (2H, m),4.06-4.20 (3H, m), 4.44-4.65 (2H, m), 5.53 (1H, d, J=5.7 Hz), 6.58 (1H,d, J=8.8 Hz), 6.90 (1H, t, J=5.4 Hz), 7.39 (1H, d, J=2.7 Hz), 7.39 (1H,dd, J=8.8, 2.7 Hz), 7.90 (1H, d, J=8.8 Hz), 7.97 (1H, d, J=8.5 Hz), 8.23(1H, d, J=8.5 Hz), 8.25 (1H, dd, J=8.8, 2.7 Hz), 8.85 (1H, br)

IR (Nujol) 3333, 1625 cm⁻¹

APCI-MS m/z 342[M+H]⁺

Synthesis Method of THK-5143

202

To a tetrahydrofuran (20 ml) solution of 201 (500 mg, 1.37 mmol), 63(1.18 g, 3.28 mmol) and triphenylphosphine (860 mg, 3.28 mmol), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (0.65ml, 3.28 mmol) was added dropwise under ice cooling and stirring,followed by stirring at the same temperature for 1 hour and furtherstirring at room temperature for 2 days. The solvent of the reactionsolution was distilled off under reduced pressure, and the residue waspurified by silica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/4) to obtain a colorless solid (1.51 g) containingthe objective product. To a chloroform (18 ml) solution of the presentproduct (1.51 g), trifluoroacetic acid (12 ml) was added dropwise underice cooling and stirring, and water (3 ml) was added, followed bystirring at room temperature for 3 days. To the reaction solution, icewater and ethyl acetate were added, and the solution was extracted withethyl acetate after adjusting the pH to 8 using an aqueous potassiumcarbonate solution. The extraction liquid was dried and the solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (eluting solvent: ethylacetate/n-hexane=2/1, ethyl acetate) and then washed with ethylacetate/n-hexane=1 to obtain 202 (520 mg, 77%) as colorless crystals.

mp 158-159° C.

APCI-MS m/z 494[M+H]⁺

Synthesis of THK-5143

To a chloroform (20 ml) suspension of 202 (510 mg, 1.03 mmol),3,4-dihydro-2H-pyran (1.87 ml, 20.7 mmol) and paratoluenesulfonic acidmonohydrate (409 mg, 2.38 mmol) were added, and the mixture was stirredat room temperature for 20 minutes. After adjusting the pH of thereaction solution to 8 by adding triethylamine, the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/1,2/1) and then recrystallized from ethyl acetate/n-hexane to obtainTHK-5143 (508 mg, 85%) as colorless crystals.

mp 93-94° C.

¹H NMR (400 MHz, DMSO-d₆) δ 1.17 (3H, t, J=7.3 Hz), 1.33-1.72 (6H, m),2.34 (3H, s), 3.30-3.37 (2H, m), 3.37-3.47 (1H, m), 3.69-3.73, 3.81-3.88(1H, m), 4.09-4.37 (5H, m), 4.71, 4.84-4.88 (1H, m), 6.58 (1H, d, J=8.8Hz), 6.91 (1H, t, J=5.3 Hz), 7.23-7.27 (1H, m), 7.29-7.31 (1H, m),7.39-7.43 (2H, m), 7.77-7.82 (2H, m), 7.98 (1H, d, J=8.8 Hz), 7.97 (1H,d, J=8.5 Hz), 8.21 (1H, d, J=8.5 Hz), 8.25 (1H, dd, J=9.1, 2.4 Hz), 8.86(1H, br)

IR (Nujol) 3347, 1622, 1602 cm⁻¹

APCI-MS m/z 578[M+H]⁺

Synthesis Method of THK-5136

Synthesis of 309

To a 1,2-dimethoxyethane (30 ml) solution of 307 (900 mg, 3.58 mmol) and308 (990 mg, 3.58 mmol), potassium carbonate (1.48 g, 10.7 mmol), water(0.62 ml) and tetrakistriphenylphosphine palladium (410 mg, 0.36 mmol)were added under an argon atmosphere, and the mixture was stirred at 80°C. for 16 hours. The reaction solution was allowed to return to roomtemperature and water was added, and the solution was extracted withethyl acetate. The extraction liquid was washed with water and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/2) to obtain 309 (1.31 g, 100%) as apale yellow solid.

mp 112-113° C.

APCI-MS m/z 366[M+H]⁺

Synthesis of 310

To a chloroform (20 ml) solution of 309 (1.31 g, 3.58 mmol),trifluoroacetic acid (2 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at the same temperature for 30minutes. To the reaction solution, ice water was added and the solutionwas extracted with ethyl acetate after adjusting the pH to 8 using anaqueous potassium carbonate solution. The extraction liquid was washedwith water and dried, and then the solvent was distilled off underreduced pressure. The residue was washed with n-hexane/ethyl acetate=4/1to obtain 310 (950 mg, 90%) as pale yellow crystals.

mp 273-274° C.

APCI-MS m/z 294[M+H]⁺

Synthesis of 312

To a tetrahydrofuran (20 ml) solution of 310 (500 mg, 1.71 mmol), 311(852 mg, 4.09 mmol) and triphenylphosphine (1.07 g, 4.09 mmol), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (0.81ml, 4.09 mmol) was added dropwise over 15 minutes under ice cooling andstirring, and the mixture was stirred at the same temperature for 1hour, followed by stirring at room temperature for 4 days. The solventof the reaction solution was distilled off under reduced pressure, andthe residue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/4, 1/2) to obtain 312 (710 mg, 86%) asa pale yellow solid.

mp 139-140° C.

APCI-MS m/z 484[M+H]⁺

Synthesis of THK-5136

To a tetrahydrofuran (20 ml) solution of 312 (700 mg, 1.45 mmol), 1Mtetra-n-butylammonium fluoride/tetrahydrofuran (1.45 ml, 1.45 mmol) wasadded dropwise at room temperature under stirring, and the mixture wasstirred at room temperature for 1 hour. To the reaction solution, waterwas added, and the solution was extracted with ethyl acetate. Theextraction liquid was dried and the solvent was distilled off underreduced pressure, and the residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/2, 2/1) andthen recrystallized from ethyl acetate to obtain THK-5136 (466 mg, 87%)as pale yellow crystals.

mp 169-170° C.

¹H NMR (400 MHz, DMSO-d₆) δ 3.12 (3H, s), 3.52 (2H, t, J=4.5 Hz),4.06-4.19 (3H, m), 4.27 (2H, t, J=4.5 Hz), 4.44-4.65 (2H, m), 5.53 (1H,d, J=4.2 Hz), 7.37-7.41 (2H, m), 7.78 (1H, d, J=1.8 Hz), 7.88-7.92 (1H,m), 8.00 (1H, d, J=8.5 Hz), 8.24 (1H, d, J=8.8 Hz), 8.56 (1H, d, J=2.1Hz)

IR (Nujol) 1620 cm⁻¹

APCI-MS m/z 370[M+H]⁺

Synthesis Method of THK-5153

Synthesis of 332

To a tetrahydrofuran (70 ml) solution of 331 (5.25 g, 35.66 mmol),N-bromosuccinimide (6.35 g, 35.66 mmol) was added at −78° C. under anargon atmosphere, and the mixture was stirred at the same temperaturefor 3 hours, followed by stirring at room temperature for 16 hours. Tothe reaction solution, water and ethyl acetate were added, and thesolution was extracted with ethyl acetate after adjusting the pH to 9using an aqueous potassium carbonate solution. The extraction liquid wasdried and the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/50) to obtain 332 (7.38 g, 91%) as acolorless oily substance.

APCI-MS m/z 226/228[M+H]⁺

Synthesis of 333

To a 1,4-dioxane (200 ml) solution of 332 (7.37 g, 32.6 mmol) and 314(9.93 g, 39 mmol), potassium acetate (9.6 g, 97.8 mmol) andPd(dppf)Cl₂.CH₂Cl₂ (1.19 g, 1.46 mmol) were added under an argonatmosphere, and the mixture was heated at reflux at 100° C. for 8 hours.The reaction solution was allowed to return to room temperature andinsolubles were filtration with celite, and the solvent of the filtratewas distilled off under reduced pressure. The residue was purified bysilica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/50) to obtain 333 (4.71 g, 53%) as a pale yellowsolid.

mp 129-130° C.

APCI-MS m/z 274[M+H]⁺

Synthesis of 334

To a 1,2-dimethoxyethane (107 ml) solution of 333 (4.46 g, 16.3 mmol)and 307 (3.28 g, 13.0 mmol), potassium carbonate (5.40 g, 39.0 mmol),water (2.27 ml) and tetrakistriphenylphosphine palladium (1.51 g, 1.3mmol) were added under an argon atmosphere, and the mixture was stirredat 80° C. for 16 hours. The reaction solution was allowed to return toroom temperature and sodium sulfate was added, followed by drying andfurther filtration with celite. The solvent of the filtrate wasdistilled off under reduced pressure and the residue was purified bysilica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/9) to obtain 334 (4.52 g, 95%) as a yellow oilysubstance.

APCI-MS m/z 363[M+H]⁺

Synthesis of 335

To a chloroform (24 ml) solution of 334 (4.51 g, 12.4 mmol),trifluoroacetic acid (16 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at the same temperature for 20minutes. To the reaction solution, water and ethyl acetate were addedand the solution was extracted with ethyl acetate after adjusting the pHto 8 using an aqueous potassium carbonate solution. The extractionliquid was washed with water and dried and the solvent was distilled offunder reduced pressure, and then the residue was washed with ethylacetate/n-hexane=1/1 to obtain 335 (3.17 g, 88%) as orange crystals.

mp 323-325° C.

APCI-MS m/z 291[M+H]⁺

Synthesis of 336

To a mixture of 335 (500 mg, 1.72 mmol), 311 (861 mg, 4.13 mmol),triphenylphosphine (1.08 g, 4.13 mmol) and tetrahydrofuran (20 ml), atetrahydrofuran (10 ml) solution of iisopropyl azodicarboxylate (0.82ml, 4.13 mmol) was added dropwise over 10 minutes under ice cooling andstirring, and the mixture was stirred at the same temperature for 1hour, followed by stirring at room temperature for 4 days. The solventof the reaction solution was distilled off under reduced pressure, andthe residue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/9) to obtain 336 (820 mg, 99%) as apale yellow solid.

mp 84-85° C.

APCI-MS m/z 481[M+H]⁺

Synthesis of THK-5153

To a tetrahydrofuran (10 ml) solution of 336 (810 mg, 1.69 mmol), 1Mtetra-n-butylammonium fluoride/tetrahydrofuran (1.69 ml, 1.69 mmol) wasadded dropwise at room temperature under stirring, and the mixture wasstirred at room temperature for 1 hour. To the reaction solution, waterwas added, and the solution was extracted with ethyl acetate. Theextraction liquid was dried and the solvent was distilled off underreduced pressure, and the residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/2) and thenrecrystallized from ethyl acetate to obtain

THK-5153 (353 mg, 57%) as pale yellow crystals.

mp 137-138° C.

¹H NMR (500 MHz, DMSO-d₆) δ 1.89-1.97 (2H, m), 2.81 (2H, t, J=6.4 Hz),2.94 (3H, s), 3.30 (2H, t, J=5.8 Hz), 4.06-4.18 (3H, m), 4.46-4.63 (2H,m), 5.55 (1H, s), 6.68 (1H, d, J=9.0 Hz), 7.38-7.43 (2H, m), 7.84 (1H,d, J=1.9 Hz), 7.91 (1H, dd, J=8.7, 2.2 Hz), 7.92-7.95 (1H, m), 7.99 (1H,d, J=8.7 Hz), 8.26 (1H, d, J=8.7 Hz)

IR (Nujol) 1598 cm⁻¹

APCI-MS m/z 367[M+H]⁺

Synthesis Method of THK-5157

Synthesis of 339

To a tetrahydrofuran (100 ml) suspension of 335 (2.00 g, 6.89 mmol), 1Mt-butoxy potassium/tetrahydrofuran (7.58 ml, 7.58 mmol) was addeddropwise and dissolved at room temperature under stirring and an argonatmosphere. The solvent of the reaction solution was distilled off underreduced pressure, and 18-crown-6 (1.82 g, 6.89 mmol) and acetonitrile(40 ml) were added to the residue. After cooling to −78° C., anacetonitrile (10 ml) solution of 338 (1.87 g, 12.4 mmol) was addeddropwise, the mixture was stirred under an argon atmosphere at roomtemperature for 16 hours. To the reaction solution, water was added andthe solution was extracted with ethyl acetate. The extraction liquid waswashed with water and dried, and then the solvent was distilled offunder reduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/4, ethylacetate) and then washed with ethyl acetate/n-hexane=1/100 to obtain 339(1.81 g, 73%) as pale yellow crystals.

mp 135-136° C.

APCI-MS m/z 361[M+H]⁺

Synthesis of 340

To a mixture of trimethylsulfoxonium iodide (1.65 g, 7.50 mmol) and DMSO(20 ml), 60% sodium hydride (300 mg, 7.50 mmol) was added under icecooling and stirring, and the mixture was stirred at room temperaturefor 30 minutes. The reaction solution was ice-cooled and atetrahydrofuran (10 ml) solution of 339 (1.80 g, 4.99 mmol) was addeddropwise, followed by stirring at room temperature for 1 hour. To thereaction solution, ice water was added, and the solution was extractedwith ethyl acetate. The extraction liquid was washed with water anddied, and then the solvent was distilled off under reduced pressure andthe residue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/4) to obtain 340 (1.74 g, 93%) as apale yellow oily substance.

APCI-MS m/z 375[M+H]⁺

Synthesis of THK-5157

A mixture of 340 (1.73 g, 4.62 mmol), n-Bu₄NH₂F₃ (232 mg, 0.46 mmol) andKHF₂ (722 mg, 9.24 mmol) was stirred at 120° C. for 2 hours. Thereaction solution was allowed to return to room temperature, water andethyl acetate were added and the solution was extracted with ethylacetate after adjusting the pH to 8 using an aqueous potassium carbonatesolution. The extraction liquid was washed with water and dried, andthen the solvent was distilled off under reduced pressure. The residuewas purified by silica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/4, 1/3) and then recrystallized from ethyl acetate toobtain THK-5157 (474 mg, 26%) as pale yellow crystals.

mp 163-164° C.

¹H NMR (400 MHz, DMSO-d₆) δ 1.21 (3H, s), 1.22 (3H, s), 1.90-1.96 (2H,m), 2.81 (2H, t, J=6.3 Hz), 2.93 (3H, s), 3.30 (2H, t, J=5.8 Hz),4.54-4.61 (1H, m), 4.67 (1H, ddd, J=48, 10, 7.1 Hz), 4.93 (1H, ddd,J=48, 10, 2.9 Hz), 4.95 (1H, d, J=6.7 Hz), 6.68 (1H, d, J=8.7 Hz),7.44-7.49 (2H, m), 7.85 (1H, d, J=2.2 Hz), 7.89-7.94 (2H, m), 7.97 (1H,d, J=8.7 Hz), 8.22 (1H, d, J=8.3 Hz)

IR (Nujol) 3267, 1620, 1599 cm⁻¹

APCI-MS m/z 395[M+H]⁺

Synthesis Method of THK-5128

Synthesis of 302

A mixture of 301 (3.65 g, 19.6 mmol), 75% methachlorobenozic acid (5.20g, 22.6 mmol) and chloroform (40 ml) was stirred at room temperature for2 hours. The reaction solution was washed in turn with an aqueous sodiumthiosulfate solution, an aqueous saturated sodium hydrogen carbonatesolution and saturated saline. The organic layer was dried and thesolvent was distilled off under reduced pressure. The residue waspurified by silica gel column chromatography (eluting solvent:chloroform/methanol=30/1, 10/1) and the washed with diisopropyl ether toobtain 302 (3.30 g, 83%) as a solid.

mp 145-146° C.

Synthesis of 303

A mixture of 302 (3.20 g, 16.3 mmol), paratoluenesulfonyl chloride (3.3g, 17.3 mmol), potassium carbonate (2.5 g, 18.1 mmol) and chloroform (60ml) was heated at reflux for 3 hours. The reaction solution was allowedto return to room temperature and water was added, and the solution wasextracted with chloroform. The extraction liquid was washed in turn withan aqueous saturated sodium hydrogen carbonate solution and a saturatedsaline and, after drying, the solvent was distilled off under reducedpressure. The residue was purified by silica gel column chromatography(eluting solvent: n-hexane/ethyl acetate=2/1) to obtain 303 (2.30 g,64%) as a colorless solid.

mp 110-111° C.

APCI-MS m/z 221[M+H]⁺

Synthesis of 305

A mixture of 303 (883 mg, 4.0 mmol), 304 (1.224 g, 4.8 mmol),1,2-dimethoxyethane (20 ml), sodium carbonate (848 mg, 8.0 mmol), water(2 ml) and tetrakistriphenylphosphine palladium (231 mg, 0.2 mmol) washeated at reflux under an argon atmosphere for 18 hours. The reactionsolution was allowed to return to room temperature and water was added,and the solution was extracted with ethyl acetate. The extraction liquidwas washed in turn with water and saturated saline and, after drying,the solvent was distilled off under reduced pressure. The residue waspurified by silica gel column chromatography (eluting solvent: ethylacetate/toluene=1/10, 1/4) to obtain 305 (1.38 g, 87%) as a colorlesssolid.

mp 160-161° C.

Synthesis of 306

To a tetrahydrofuran (30 ml) suspension of NaBH₄ (380 mg, 10 mmol), atetrahydrofuran (5 ml) solution of BF₃.Et₂O (1.86 g, 13.1 mmol) wasadded dropwise at 0° C., and the mixture was stirred at the sametemperature for 30 minutes. To the present reaction solution, atetrahydrofuran (10 ml) solution of 305 (1.30 g, 3.28 mmol) was addeddropwise at 0 to 5° C., and the solution was stirred at the sametemperature for 20 minutes, followed by stirring at room temperature for1 hour. The reaction solution was ice-cooled and an aqueous saturatedsodium hydrogen carbonate solution was added dropwise, and the solutionwas stirred and then extracted with ethyl acetate. The extraction liquidwas washed in turn with water and saturated saline and, after drying,the solvent was distilled off under reduced pressure. The residue waspurified by silica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/4, 1/3) and then washed with n-hexane-diisopropylether to obtain 306 (980 mg, 78%) as an orange solid.

mp 175-176° C.

APCI-MS m/z 382[M+H]⁺

Synthesis of THK-5128

To a methylene chloride (15 ml) solution of 306 (900 mg, 2.36 mmol),trifluoroacetic acid (3 ml) was added dropwise under ice cooling andstirring, and the mixture was stirred at room temperature for 1 hour.The reaction solution was ice-cooled and diluted with ethyl acetate, andthe solution was made basic with 28% ammonia water, followed by liquidseparation. The organic layer was washed in turn with water andsaturated saline and, after drying, the solvent was distilled off underreduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: toluene, ethyl acetate/toluene=1/3) andthen recrystallized from ethyl acetate to obtain THK-5128 (545 mg, 82%)as yellow crystals.

mp 204-205° C. (dec.)

¹H NMR (400 MHz, CDCl₃) δ 3.08 (6H, s), 3.86 (2H, brs), 6.80 (1H, d,J=2.7 Hz), 6.87 (1H, dd, J=9.2, 8.8 Hz), 7.36 (1H, dd, J=9.4, 2.7 Hz),7.66 (1H, d, J=8.8 Hz), 7.72 (1H, dd, J=8.8, 1.8 Hz), 7.86 (1H, dd,J=13, 1.8 Hz), 7.96 (1H, d, J=8.8 Hz), 7.97 (1H, d, J=9.4 Hz).

IR (Nujol) 3460, 3304, 3181, 1645, 1619, 1589 cm⁻¹

APCI-MS m/z 282[M+H]⁺

Synthesis of THK-5147

Synthesis of 315

A mixture of 313 (18.76 g, 0.1 mol), 314 (27.93 g, 0.11 mol), potassiumacetate (14.72 g, 0.15 mol), Pd₂(dba)₃ (2.75 g, 3 mmol),tricyclohexylphosphine (2.02 g, 7.2 mmol) and 1,4-dioxane (200 ml) wasstirred under an argon atmosphere at 90° C. for 16 hours. The reactionsolution was allowed to return to room temperature and poured into ethylacetate-water, and then the solution was extracted with ethyl acetate.The extraction liquid was washed with saturated saline and dried, andthen the solvent was distilled off under reduced pressure. The residuewas purified by silica gel flash column chromatography (eluting solvent:n-hexane→n-hexane/ethyl acetate=19/1, 9/1, 6/1, 4/1) to obtain 315(26.74 g, 96%) as a pale yellow solid.

APCI-MS m/z 280[M+H]⁺

Synthesis of 317

To a pyridine (1 L) suspension of 316 (29.51 g, 146 mmol),trifluoromethanesulfonic anhydride (48 ml, 292 mmol) was added dropwiseat −30 to −20° C., and the mixture was stirred at 10° C. for 1 hour. Tothe reaction solution, ice water was added after distilling off avolatile substance at 40° C. or lower, the solution was extracted withethyl acetate. The extraction liquid was washed with saturated salineand dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel flash columnchromatography (eluting solvent: n-hexane→n-hexane/ethyl acetate=2/1,1/1, 1/2, 1/5) and then recrystallized from ethyl acetate-diisopropylether to obtain 317 (30.86 g, 63%) as pale pink crystals.

APCI-MS m/z 335[M+H]⁺

Synthesis of 318

A mixture of 317 (29.11 g, 87 mmol), 315 (26.74 g, 96 mmol), an aqueous2M sodium carbonate solution (100 ml), tetrakistriphenylphosphinepalladium (5 g, 4.3 mmol) and 1,2-dimethoxyethane (300 ml) was stirredunder an argon atmosphere at 90° C. for 16 hours. The reaction solutionwas allowed to return to room temperature and 1,2-dimethoxyethane wasdistilled off under reduced pressure, and chloroform/methanol (=1/1) wasadded and then insolubles were removed by filtration. The filtrate waswashed with saturated saline and dried the solvent was distilled offunder reduced pressure. The residue was purified by silica gel flashcolumn chromatography (eluting solvent: chloroform/tetrahydrofuran=4/1)and then washed with chloroform-diisopropyl ether to obtain 318 (20.5 g,76%) as a yellow solid.

APCI-MS m/z 338[M+H]⁺

Synthesis of 319

A mixture of 318 (1.0 g, 29.6 mmol), lithium chloride (1.26 g, 29.7mmol) and hexamethylphosphoric triamide (5 ml) was stirred at 110° C.

for 2 days. The reaction solution was allowed to return to roomtemperature and water was added. The solution was acidified with 10%hydrochloric acid water and made basic with an aqueous saturated sodiumhydrogen carbonate solution, and then extracted with ethylacetate-tetrahydrofuran. The extraction liquid was washed with saturatedsaline and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel flash columnchromatography (eluting solvent: chloroform/tetrahydrofuran=4/1, 3/2)and then recrystallized from tetrahydrofuran-diisopropyl ether to obtain319 (745 mg, 78%) as a yellow solid.

APCI-MS m/z 324[M+H]⁺

Synthesis of 320

To a mixture of 319 (323 mg, 1.0 mmol), 311 (208 mg, 1.0 mmol),triphenylphosphine (394 mg, 1.5 mmol) and tetrahydrofuran (5 ml), atetrahydrofuran (2 ml) solution of diisopropyl azodicarboxylate (300 mg,1.5 mmol) was added dropwise under ice cooling and stirring, followed bystirring at the same temperature for 30 minutes and further stirring atroom temperature for 20 hours. The reaction solution was purified bysilica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/1) to obtain 320 (403 mg, 78%) as a yellow caramel.

APCI-MS m/z 514[M+H]⁺

Synthesis of 321

To a N,N-dimethylformamide (20 ml) suspension of 60% sodium hydride (345mg, 8.6 mmol), an N,N-dimethylformamide (10 ml) solution of 320 (3.70 g,7.2 mmol) was added dropwise under an argon atmosphere, and the mixturewas stirred at 0 to 5° C., followed by stirring at −15° C. for 30minutes. The reaction solution was allowed to return to 0 to 5° C. andan N, N-dimethylformamide (2 ml) solution of methyl iodide (2.2 g, 15mmol) was added dropwise, and the solution was stirred at roomtemperature for 1 hour. The reaction solution was poured into ice water,and the solution was extracted with ethyl acetate. The extraction liquidwas washed with saturated saline and dried, and then the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel flash column chromatography (eluting solvent: ethylacetate/n-hexane=1/1) to obtain 321 (3.30 g, 89%) as a pale yellowsolid.

mp 97-99° C.

APCI-MS m/z 528[M+H]⁺

Synthesis of 322

A mixture of 321 (2.18 g, 4.13 mmol) and 48% HBr (25 ml) was stirred at90 to 95° C. for 2 hours. The reaction solution was cooled and was madebasic with 28% ammonia water, and then extracted with ethyl acetate. Theextraction liquid was washed with saturated saline and dried, and thenthe solvent was distilled off under reduced pressure. The residue waswashed with diisopropyl ether to obtain 322 (1.45 g, 95%) as an orangesolid.

mp 130-132° C.

IR (Nujol) 3379, 1624, 1605 cm⁻¹

APCI-MS m/z 372[M+H]⁺

Synthesis of THK-5147

A mixture of 322 (250 mg, 0.673 mmol), 5 nCl₂.2H₂O (450 mg, 1.99 mmol)and ethanol (10 ml) was heated at reflux for 1 hour. The reactionsolution was allowed to return to room temperature, diluted with ethylacetate and then made basic with 28% ammonia water. Insolubles wereremoved by filtration with celite, and the organic layer of the filtratewas separated and washed with saturated saline. After drying, thesolvent was distilled off under reduced pressure. The residue waspurified by NH silica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/4) to obtain THK-5147 (175 mg, 76%) as a yellowsolid.

mp 122-124° C.

¹H NMR (400 MHz, CDCl₃) δ 2.95 (3H, s), 3.96 (3H, br), 4.23-4.37 (3H,m), 4.49-4.61 (1H, m), 4.62-4.73 (1H, m), 6.69 (1H, d, J=2.7 Hz), 6.81(1H, d, J=8.2 Hz), 7.08 (1H, dd, J=9.0, 2.7 Hz), 7.53 (1H, dd, J=8.2,1.8 Hz), 7.68 (1H, d, J=8.8 Hz), 7.74 (1H, d, J=1.8 Hz), 7.91 (1H, d,J=9.0 Hz), 7.94 (1H, d, J=8.8 Hz)

IR (Nujol) 3429, 3342, 1625, 1595 cm⁻¹

APCI-MS m/z 342[M+H]⁺

Synthesis Method of THK-5148

Synthesis of 325

Under an argon atmosphere, a tetrahydrofuran (15 ml) solution of 324(3.1 g, 36 mmol) was added dropwise to a tetrahydrofuran (30 ml)suspension of 60% sodium hydride (1.44 g, 36 mmol) at room temperature,and the mixture was heated at reflux for 30 minutes. The reactionsolution was cooled to 0 to 5° C. and a tetrahydrofuran (15 ml) solutionof 323 (7.2 g, 33 mmol) was added dropwise, and the solution was stirredat room temperature for 16 hours. The reaction solution was poured intoa cooled aqueous ammonium chloride solution, and then the solution wasextracted with ethyl acetate. The extraction liquid was dried and thesolvent was distilled off under reduced pressure, and then the residuewas purified by silica gel flash column chromatography (elutingsolvent:n-hexane→ethyl acetate/n-hexane=1/50, 1/20) to obtain 325 (8.46g, 90%) as a yellow oily substance.

APCI-MS m/z 303/305[M+NE₄]⁺

Synthesis of 326

A mixture of 325 (7.0 g, 24.5 mmol), 75% methachlorobenozic acid (20.26g, 88.1 mmol) and chloroform (70 ml) was stirred at room temperature for2 days. The reaction solution was ice-cooled and an aqueous saturatedsodium hydrogen carbonate solution was added, and the solution wasextracted with chloroform. The extraction liquid was dried and thesolvent was distilled off under reduced pressure, and then the residuewas purified by silica gel flash column chromatography (eluting solvent:n-hexane→ethyl acetate/n-hexane=1/20, 1/10) to obtain 326 (4.197 g, 57%)as a yellow oily substance.

APCI-MS m/z 319/321[M+NH₄]⁺

Synthesis of 327

A mixture of 326 (4.119 g, 13.6 mmol), n-Bu₄NH₂F₃ (410 mg, 1.36 mmol)and KHF₂ (2.13 g, 27.27 mmol) was stirred at 120° C. for 6 hours. Thereaction solution was allowed to return to room temperature andchloroform was added, and then insolubles were removed by filtration.The filtrate was dried and the solvent was distilled off under reducedpressure, and then the residue was purified by silica gel flash columnchromatography (eluting solvent: n-hexane→ethyl acetate/n-hexane=1/20,1/10, 1/6, 1/4) to obtain 327 (1.3 g, 30%) as a yellow solid.

APCI-MS m/z 339/341[M+NH₄]⁺

Synthesis of 328

A mixture of 327 (1.29 g, 4.0 mmol), 314 (1.06 g, 4.16 mmol), potassiumacetate (1.18 g, 12 mmol), Pd(dppf)Cl₂ CH₂Cl₂ (229 mg, 0.28 mmol) and1,4-dioxane (20 ml) was heated at reflux for 16 hours under an argonatmosphere. The reaction solution was allowed to return to roomtemperature and the solvent was distilled off under reduced pressure. Tothe residue, ethyl acetate and an aqueous saturated sodium hydrogencarbonate solution were added, and then the solution was extracted withethyl acetate. The extraction liquid was dried and the solvent wasdistilled off under reduced pressure, and then the residue was purifiedby silica gel flash column chromatography (eluting solvent:n-hexane→ethyl acetate/n-hexane=1/10, 1/4, 1/2) to obtain 328 (1.438 g,97%) as a brown oily substance.

APCI-MS m/z 387[M+NH₄]⁺

Synthesis of 330

A mixture of 328 (1.4 g, 3.8 mmol), 329 (729 mg, 3.5 mmol), an aqueous2M sodium carbonate solution (3.5 ml), tetrakistriphenylphosphinepalladium (202 mg, 0.175 mmol) and 1,2-dimethoxyethane (20 ml) wasstirred at 90° C. for 5 hours under an argon atmosphere. The reactionsolution was allowed to return to room temperature and1,2-dimethoxyethane was distilled off under reduced pressure. Chloroformwas added and insolubles were removed by filtration, followed by washingwith chloroform/methanol (=9/1). The filtrate and the wash werecombined, and the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel flash column chromatography (elutingsolvent: n-hexane→ethyl acetate/n-hexane=1/10, 1/4, 1/2) and then washedwith n-hexane to obtain 330 (1.448 g, 99%) as a yellow solid.

APCI-MS m/z 416[M+H]⁺

Synthesis of THK-5148

330 (1.44 g, 3.47 mmol), 10% Pd—C (300 mg) and ethanol-methanol (60ml-20 ml) were stirred under hydrogen pressure (40 psi) at roomtemperature for 16 hours. The catalyst was removed by filtration andPd—C (150 mg) was added to the filtrate, followed by further stirringunder hydrogen pressure (40 psi) at room temperature for 1.5 hours. Thecatalyst was removed by filtration and the solvent of the filtrate wasdistilled off under reduced pressure. To the residue, methanol (50ml)-acetic acid (5 ml), an aqueous 36% formaldehyde solution (3.1 g,37.2 mmol) and a picoline borane complex (1.5 g, 14 mmol) were added,followed by stirring at room temperature for 16 hours. The solvent ofthe reaction solution was distilled off under reduced pressure and anaqueous saturated sodium hydrogen carbonate solution the residue, andthe solution was extracted with chloroform. The extraction liquid wasdried and the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel flash column chromatography (elutingsolvent: chloroform→chloroform/methanol=50/1) and then recrystallizedfrom chloroform-n-hexane to obtain THK-5148 (832 mg, 58%) as a yellowishorange solid.

mp 182-184° C.

¹H NMR (400 MHz, DMSO-d₆) δ 1.25, 1.23 (6H, each s), 2.05 (6H, s), 2.82(6H, s), 4.45-4.51 (1H, m), 4.81 (1H, ddd, J=48, 10, 6.5 Hz), 4.90 (1H,ddd, J=46, 10, 2.4 Hz), 5.09 (1H, brs), 6.94 (1H, d, J=2.7 Hz), 7.02(1H, d, J=8.4 Hz), 7.44 (1H, dd, J=9.4, 2.7 Hz), 7.73 (1H, dd, J=8.5,1.8 Hz), 7.85 (1H, d, J=9.4 Hz), 7.89 (1H, d, J=8.7 Hz), 7.91 (1H, d,J=2.1 Hz), 8.13 (1H, d, J=8.8 Hz)

IR (Nujol) 1819, 1375 cm⁻¹

APCI-MS m/z 412[M+H]⁺

Synthesis Method of THK-5155

Synthesis of 337

320 (1.18 g, 2.3 mmol), 10% Pd—C (moisture 50%; 250 mg) and ethanol (30ml) were stirred under hydrogen pressure (40 psi) at room temperaturefor 16 hours. The catalyst was removed by filtration and the solvent ofthe filtrate was distilled off under reduced pressure, and then theresidue was purified by NH silica gel flash column chromatography(eluting solvent: chloroform) to obtain 337 (1.03 g, 93%) as a yellowfoam-like substance.

Synthesis of THK-5155

A mixture of 337 (1.025 g, 2.12 mmol) and 48% HBr (10 ml) was stirred at110° C. for 5 hours. The reaction solution was allowed to return to roomtemperature and the solvent was distilled off under reduced pressure. Tothe residue, an aqueous saturated sodium hydrogen carbonate solution wasadded, and the solution was extracted with ethylacetate-tetrahydrofuran. The extraction liquid was dried and the solventwas distilled off under reduced pressure, and then the residue waspurified by NH silica gel flash column chromatography (eluting solvent:chloroform/methanol=50/1) to obtain a pale yellow amorphous (684 mg).The present product was converted into a hydrochloride by a conventionalmethod using 4M hydrochloric acid/ethyl acetate to obtain THK-5155 (670mg, 79%) as an orange solid.

mp 232-235° C.

¹H NMR (400 MHz, DMSO-d₆) δ 4.05-4.25 (3H, m), 4.50-4.70 (2H, m), 6.94(1H, d, J=8.2 Hz), 7.18 (1H, brs), 7.49 (1H, dd, J=9.0, 2.4 Hz), 7.72(1H, dd, J=8.4, 1.8 Hz), 7.81 (1H, d, J=1.8 Hz), 8.19 (1H, d, J=9.0 Hz),8.39 (1H, d, J=9.0 Hz), 8.62 (1H, d, J=9.0 Hz)

IR (Nujol) 1624, 1460 cm⁻¹

APCI-MS m/z 328[M+H]⁺

Synthesis Method of THK-5156

Synthesis of THK-5156

To a mixture of THK-5155 (free form) (450 mg, 1.4 mmol), an aqueous 36%formaldehyde solution (0.97 g, 11.6 mmol) and methanol (10 ml)-aceticacid (1 ml), a picoline borane complex (441 mg, 4.1 mmol) was addedlittle by little, and the mixture was stirred at room temperature for 16hours. The solvent of the reaction solution was distilled off underreduced pressure and 10% hydrochloric acid water was added to theresidue, and the solution was stirred at room temperature for 10minutes. The reaction solution was extracted with chloroform after beingmade basic with an aqueous potassium carbonate solution. The extractionliquid was dried and the solvent was distilled off under reducedpressure, and then the residue was purified by silica gel flash columnchromatography (eluting solvent: chloroformchloroform/tetrahydrofuran=9/1, 6/1, 4/1) to obtain THK-5156 (328 mg,62%) as an orange solid.

mp 187-188.5° C.

¹H NMR (400 MHz, DMSO-d₆) δ 2.82 (6H, s), 3.05 (6H, s), 4.10-4.20 (3H,m), 4.40-4.70 (2H, m), 5.50 (1H, brs), 6.95 (1H, d, J=2.4 Hz), 6.98 (1H,d, J=8.5 Hz), 7.45 (1H, dd, J=9.0, 2.4 Hz), 7.72 (1H, dd, J=8.4, 1.8Hz), 7.78 (1H, d, J=1.8 Hz), 7.87 (1H, d, J=9.4 Hz), 7.95 (1H, d, J=8.8Hz), 8.14 (1H, d, J=8.8 Hz)

IR (Nujol) 1620, 1463, 1375 cm⁻¹

APCI-MS m/z 384[M+H]⁺

Synthesis Method of THK-5158

Synthesis of 342

To a 1,2-dimethoxyethane (364 ml) solution of 341 (9.14 g, 44.23 mmol)and 315 (14.57 g, 52.20 mmol), potassium carbonate (18.34 g, 133 mmol),water (7.7 ml) and tetrakistriphenylphosphine palladium (5.11 g, 4.42mmol) were added under an argon atmosphere, and the mixture was stirredat 80° C. for 16 hours. The reaction solution was allowed to return toroom temperature and ethyl acetate was added, and then insolubles wereremoved by filtration with celite and the solution was washed withchloroform/methanol (=1/1). The filtrate and the wash were combined, andthe solvent was distilled off under reduced pressure. The residue waspurified by silica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/1, 3/1) and then washed with ethyl acetate to obtain342 (9.55 g, 67%) as orange crystals.

mp 216-217° C.

APCI-MS m/z 324[M+H]⁺

Synthesis of 343

A mixture of 342 (6.5 g, 20.1 mmol), lithium chloride (8.48 g, 0.2 mol)and hexamethylphosphoric triamide (65 ml) was stirred at 110° C. for 41hour. The reaction solution was allowed to return to room temperatureand poured into an aqueous citric acid solution, and the solution wasstirred. The precipitate was collected by filtration, washed with waterand then dissolved in tetrahydrofuran. The tetrahydrofuran solution wasdried and the solvent was distilled off under reduced pressure, and thenthe residue was washed with ethyl acetate to obtain 343 (5.74 g, 92%) asa brown solid.

mp 222-223° C.

Synthesis of 344

To a tetrahydrofuran (110 ml) solution of 343 (5.73 g, 18.5 mmol), 311(5.65 g, 27.1 mmol) and triphenylphosphine (8.26 g, 31.5 mmol), atetrahydrofuran (20 ml) solution of diisopropyl azodicarboxylate (6.37g, 31.5 mmol) was added dropwise at −5 to 0° C. over 20 minutes, and themixture was stirred at the same temperature for 30 minutes, followed bystirring at room temperature for 66 hours. To the reaction solution, 311(565 mg, 2.7 mmol), triphenylphosphine (826 mg, 3.15 mmol) anddiisopropyl azodicarboxylate (638 mg, 3.15 mmol) were added, and thesolution was further stirred at room temperature for 6 hours. Thesolvent of the reaction solution was distilled off under reducedpressure and the residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/2, 1/1) toobtain 344 (7.67 g, 83%) as a solid.

mp 150-152° C.

APCI-MS m/z 500[M+H]⁺

Synthesis of 345

344 (1.0 g, 2 mmol), 10% Pd—C (150 mg) and ethanol (20 ml) were stirredunder hydrogen pressure (40 psi) at room temperature for 16 hours. Thecatalyst was removed by filtration and the solvent was distilled offunder reduced pressure. The pale yellow amorphous as the residue wasdissolved by adding ethyl formate (20 ml) and the solution was heated atreflux for 16 hours. The reaction solution was allowed to return to roomtemperature and the solvent was distilled off under reduced pressure.The residue was purified by silica gel flash column chromatography(eluting solvent: tetrahydrofuran/chloroform=1/19, 1/9, 1/4, 1/3, 1/1,methanol/chloroform=1/9, 1/1) and then washed with ethylacetate/n-hexane to obtain 345 (860 mg, 86%) as an orange solid.

APCI-MS m/z 498[M+H]⁺

Synthesis of THK-5158

To a tetrahydrofuran (10 ml) suspension of NaBH₄ (392 mg, 10.4 mmol), atetrahydrofuran (10 ml) solution of BF₃ Et₂O (1.96 g, 13.8 mmol) wasadded dropwise at 0° C., and the mixture was stirred at room temperaturefor 1 hour. To the present reaction solution, a tetrahydrofuran (10 ml)solution of 345 (855 mg, 1.7 mmol) was added dropwise at 0 to 5° C., andthe solution was stirred at room temperature for 2 hours, followed bystirring under heating at reflux for 3 hours. The reaction solution wasallowed to return to room temperature and the solvent was distilled offunder reduced pressure. To the residue, 10% hydrochloric acid water wasadded, followed by stirring at room temperature for 1 hour. The reactionsolution was made basic by adding potassium carbonate and then extractedwith chloroform. The extraction liquid was dried and the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel flash column chromatography (eluting solvent:chloroform→tetrahydrofuran/chloroform=1/19, 1/9, 1/4), purified again bysilica gel flash column chromatography (eluting solvent: n-hexane→ethylacetate/n-hexane=1/1, 6/4) and then washed with diisopropyl ether andthen n-hexane to obtain THK-5158 (331 mg, 54%) as an orange solid.

mp 150-152° C.

¹H NMR (400 MHz, DMSO-d₆) δ 2.79 (3H, d, J=5.8 Hz), 2.80 (3H, d, J=5.1Hz), 3.95-4.20 (3H, m), 4.50-4.70 (2H, m), 5.47 (1H, d, J=5.8 Hz), 5.53(1H, d, J=5.1 Hz), 6.12 (1H, d, J=5.1 Hz), 6.56 (1H, d, J=8.8 Hz), 6.62(1H, d, J=2.7 Hz), 7.13 (1H, dd, J=9.1, 2.4 Hz), 7.64-7.68 (2H, m), 7.70(1H, d, J=9.1 Hz), 7.84 (1H, d, J=8.8 Hz), 7.99 (1H, d, J=8.7 Hz)

IR (Nujol) 1621 cm⁻¹

APCI-MS m/z 356[M+H]⁺

Synthesis Method of THK-5159

Synthesis of 353

A mixture of 320 (100 mg, 0.195 mmol) and 48% HBr (1 ml) was stirred at90 to 95° C. for 1 hour. The reaction solution was allowed to return toroom temperature and made basic concentrated ammonia water, followed byextraction with ethyl acetate. The extraction liquid was washed withsaturated saline and dried, and then the solvent was distilled off underreduced pressure. The residue was washed with ethyl acetate-diisopropylether to obtain 353 (45 mg, 65%) as an orange solid.

mp 153-154° C.

APCI-MS m/z 358[M+H]⁺

Synthesis of 354

To a mixture of 353 (167 mg, 0.467 mmol), an aqueous 36% formaldehydesolution (1 ml) and ethanol (5 ml)-acetic acid (0.5 ml), a picolineborane complex (150 mg, 1.4 mmol) was added little by little, followedby stirring at room temperature for 3 hours. The reaction solution wasdiluted with ethyl acetate, washed in turn with an aqueous saturatedsodium hydrogen carbonate solution and saturated saline and dried, andthen the solvent was distilled off under reduced pressure. The residuewas purified by silica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/2) and then washed with ethyl acetate/n-hexane (=1/3)to obtain 354 (161 mg, 89%) as a brown solid.

mp 169-171° C.

Synthesis of THK-5159

A mixture of 354 (270 mg, 0.7 mmol), 10% Pd—C (moisture of about 50%;100 mg), ammonium formate (440 mg, 7 mmol) and methanol (10ml)-tetrahydrofuran (5 ml) was stirred under an argon atmosphere at roomtemperature for 3 hours. The reaction solution was diluted with ethylacetate and insolubles were removed by filtration. The filtrate waswashed in turn with water and saturated saline and dried, and then thesolvent was distilled off under reduced pressure. The residue waspurified by NH silica gel flash column chromatography (eluting solvent:n-hexane/ethyl acetate=1/1, 1/2, 1/3, 1/4, 1/5) and then washed withn-hexane/diisopropyl ether to obtain THK-5159 (216 mg, 87%) as a palebrown solid.

mp 169-170° C.

¹H NMR (400 MHz, CDCl₃) δ 2.94 (1H, br), 3.08 (6H, s), 3.96 (2H, br),4.24-4.36 (3H, m), 4.52-4.61 (1H, m), 4.64-4.73 (1H, m), 6.81 (1H, d,J=3.0 Hz), 6.81 (1H, d, J=8.0 Hz), 7.36 (1H, dd, J=9.4, 3.0 Hz), 7.53(1H, dd, J=8.0, 1.2 Hz), 7.68 (1H, d, J=8.8 Hz), 7.76 (1H, br), 7.96(1H, d, J=8.8 Hz), 7.99 (1H, d, J=9.4 Hz)

IR (Nujol) 3345, 1621, 1590 cm⁻¹

APCI-MS m/z 356[M+H]⁺

Synthesis Method of THK-5160

Synthesis of 355

344 (2.65 g, 5.3 mmol), 10% Pd—C (moisture of about 50%; 350 mg) andethyl acetate (40 ml)-tetrahydrofuran (20 ml) were stirred under ahydrogen atmosphere at room temperature for 21 hour. The catalyst wasremoved by filtration and the solvent of the filtrate was distilled offunder reduced pressure. The residue was dissolved in ethyl acetate (50ml) and 10% Pd—C (moisture of about 50%; 300 mg) was added, and thesolution was stirred under a hydrogen atmosphere at room temperature for2 days. The catalyst was removed by filtration and the solvent of thefiltrate was distilled off under reduced pressure, and the residue waspurified by silica gel column chromatography (eluting solvent:n-hexane/ethyl acetate=1/1, 2/3, ethyl acetate) to obtain 355 (2.27 g,91%).

APCI-MS m/z 470[M+H]⁺

Synthesis of 356

To a mixture of 355 (1.20 g, 2.55 mmol), an aqueous 35% formaldehydesolution (2.2 g, 26 mmol) and ethanol (20 ml)-acetic acid (2 ml), apicoline borane complex (545 mg, 5.1 mmol) was added little by little,followed by stirring at room temperature for 6 hours. To the reactionsolution, an aqueous 35% formaldehyde solution (2.2 g, 26 mmol) and apicoline borane complex (545 mg, 5.1 mmol) were added, followed byfurther stirring at room temperature for 2 hours. The reaction solutionwas diluted with ethyl acetate, washed in turn with water, diluteammonia water and saturated saline and dried, and then the solvent wasdistilled off under reduced pressure. The residue was washed withdiisopropyl ether to obtain 356 (922 mg, 73%) as a pale brown solid.

mp 164-166° C.

APCI-MS m/z 498 [M+H]⁺

Synthesis of THK-5160

A mixture of 356 (350 mg, 0.7 mmol) and 48% HBr (3 ml) was stirred at 90to 95° C. for 1 hour. The reaction solution was allowed to return toroom temperature and diluted with ethyl acetate and ice water was added.Then, the solution was made basic with concentrated ammonia water andextracted with ethyl acetate. The extraction liquid was washed in turnwith water and saturated saline and dried, and then the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel flash column chromatography (eluting solvent: n-hexane/ethylacetate=1/2, 1/4, ethyl acetate) to obtain THK-5160 (179 mg, 72%) as abeige solid.

mp 158-160° C.

¹H NMR (400 MHz, CDCl₃) δ 2.85 (6H, s), 3.95 (2H, brs), 4.04-4.15 (1H,m), 4.24-4.35 (2H, m), 4.43-4.52 (1H, m), 4.55-4.63 (1H, m), 5.20 (1H,br), 6.92 (1H, d, J=2.4 Hz), 7.11 (1H, d, J=8.5 Hz), 7.17 (1H, dd,J=8.5, 2.7 Hz), 7.71 (1H, d, J=8.8 Hz), 7.72 (1H, dd, J=8.5, 2.1 Hz),7.86 (1H, d, J=2.1 Hz), 7.94 (1H, d, J=8.5 Hz), 7.95 (1H, d, J=8.8 Hz)

IR (Nujol) 3462, 3345, 1633 cm⁻¹

APCI-MS m/z 356[M+H]⁺

Synthesis Method of THK-5161

Synthesis of 357

Under an argon atmosphere, a tetrahydrofuran (5 ml) solution of BF₃.Et₂O(1.23 g, 8.7 mmol) was added dropwise to a tetrahydrofuran (10 ml)suspension of NaBH₄ (252 mg, 6.6 mmol) at 0° C., and the mixture wasstirred at the same temperature for 20 minutes. To the present reactionsolution, a tetrahydrofuran (3 ml) solution of 356 (630 mg, 1.27 mmol)was added dropwise at 0 to 5° C., followed by stirring at 5° C. for 1hour. To the reaction solution, ice water and ethyl acetate were added,and the solution was made basic with concentrated ammonia water and thenextracted with ethyl acetate. The extraction liquid was washed in turnwith water and then saturated saline, and dried and then the solvent wasdistilled off under reduced pressure. The residue was purified by NHsilica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/8, 1/6) to obtain 357 (440 mg, 72%) as a pale yellowsolid.

APCI-MS m/z 484[M+H]⁺

Synthesis of THK-5161

A mixture of 357 (440 mg, 0.91 mmol) and 48% HBr (3 ml) was stirred at50° C. for 10 minutes. The reaction solution was made basic withconcentrated ammonia water under ice cooling, and then extracted withethyl acetate. The extraction liquid was washed in turn with water andthen saturated saline, and dried the solvent was distilled off underreduced pressure. The residue was purified by NH silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=2/1, 3/1) toobtain THK-5161 (280 mg, 83%) as a pale yellow solid.

mp 140-141° C.

¹H NMR (400 MHz, CDCl₃) δ 2.84 (6H, s), 2.96 (3H, brs), 4.00-4.15 (2H,m), 4.24-4.35 (2H, m), 4.43-4.51 (1H, m), 4.55-4.63 (1H, m), 5.26 (1H,br), 6.71 (1H, d, J=2.7 Hz), 7.10 (1H, d, J=9.0 Hz), 7.10 (1H, dd,J=9.0, 2.7 Hz), 7.71 (1H, d, J=8.5 Hz), 7.72 (1H, dd, J=8.5, 2.0 Hz),7.86 (1H, d, J=2.0 Hz), 7.92 (1H, d, J=9.0 Hz), 7.98 (1H, d, J=8.8 Hz)

IR (Nujol) 3462, 1627 cm⁻¹

APCI-MS m/z 370[M+H]⁺

Synthesis Method of THK-5162

Synthesis of 381

To an N,N-dimethylformamide (30 ml) suspension of 361 (900 mg, 2.91mmol) and 380 (4.00 g, 8.7 mmol), potassium carbonate (1.21 g, 8.7 mmol)was added at room temperature under stirring, and the mixture wasstirred at room temperature for 4 days. The reaction solution wasextracted with ethyl acetate after adding water and ethyl acetate. Theextraction liquid was washed with water and dried, and then the solventwas distilled off under reduced pressure. The residue was purified bysilica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/2, 1/1) to obtain 381 (1.73 g, 100%) as a red oilysubstance.

APCI-MS m/z 596 [M+H]+

Synthesis of 382

A mixture of 381 (1.73 g, 2.9 mmol) and 1M tetra-n-butylammoniumfluoride/tetrahydrofuran (14.6 ml, 14.6 mmol) was stirred at roomtemperature for 16 hours. The reaction solution was diluted with ethylacetate, washed with water and dried, and then the solvent was distilledoff under reduced pressure. The residue was purified by silica gelcolumn chromatography (eluting solvent: ethyl acetate/n-hexane=1/2, 2/3)to obtain 382 (960 mg, 74%) as a red resinous substance.

APCI-MS m/z 444[M+H]⁺

Synthesis of 383

A mixture of 382 (950 mg, 2.14 mmol), 10% Pd—C (moisture of about 50%;190 mg), ammonium formate (1.35 g, 21.4 mmol) and methanol (20ml)-tetrahydrofuran (10 ml) was stirred under an argon atmosphere atroom temperature for 1 hour. The reaction solution was diluted withethyl acetate and insolubles were removed by filtration, and then thesolvent of the filtrate was distilled off under reduced pressure. Theresidue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/2, 2/1) to obtain 383 (850 mg, 96%) asa yellow resinous substance.

APCI-MS m/z 414[M+H]⁺

Synthesis of THK-5162

A mixture of 383 (600 mg, 1.45 mmol), an aqueous 35% formaldehydesolution (373 mg, 4.35 mmol), magnesium sulfate (7 g), tetrahydrofuran(40 ml) and isopropanol (20 ml) was stirred at room temperature for 3hours. To the reaction solution, NaBH₄ (274 mg, 7.2 mmol) was added, andthe solution was stirred at room temperature for 3 days. Insolubles wereremoved by filtration from the reaction solution, and the solvent of thefiltrate was distilled off under reduced pressure. The residue waspurified by silica gel column chromatography (eluting solvent: ethylacetate/n-hexane=2/1) and the obtained orange resinous substance (580mg; free form) was converted into a hydrochloride by treating with 4Mhydrochloric acid/ethyl acetate, and then recrystallized from ethanol toobtain THK-5162 (516 mg, 71%) as orange crystals.

mp 197-198° C.

¹H NMR (400 MHz, DMSO-d₆) δ 2.87 (3H, s), 3.10 (6H, s), 3.56-3.64 (3H,m), 3.64-3.68 (2H, m), 3.69-3.72 (1H, m), 3.84-3.88 (2H, m), 4.38 (2H,dd, J=5.4, 3.9 Hz), 4.53 (2H, dt, J=48, 4.3 Hz), 6.72 (1H, d, J=8.8 Hz),7.18 (1H, d, J=2.7 Hz), 7.68 (1H, dd, J=9.5, 2.9 Hz), 7.84 (1H, dd,J=8.5, 2.1 Hz), 7.90 (1H, d, J=2.1 Hz), 8.24 (1H, d, J=9.1 Hz), 8.58(1H, d, J=9.4 Hz), 8.63 (1H, d, J=9.1 Hz)

IR (Nujol) 3376, 2582, 1643, 1603 cm⁻¹

APCI-MS m/z 428[M+H]⁺

Synthesis Method of THK-5163

Synthesis of 359

To a mixture of 358 (2.20 g, 12.3 mmol), an aqueous 36% formaldehydesolution (20.5 g, 246 mmol) and methanol (250 ml)-acetic acid (20 ml), apicoline borane complex (7.91 g, 73.95 mmol) was added little by little,followed by stirring at room temperature for 16 hours. To the reactionsolution, ethyl acetate-water was added and the solution was extractedwith ethyl acetate after adjusting the pH to 8 using an aqueouspotassium carbonate solution. The extraction liquid was washed withwater and dried the solvent was distilled off under reduced pressure andthe residue was purified by silica gel column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/4) to obtain 359 (2.54 g, 99%) as apale yellow solid.

mp 74-75° C.

APCI-MS m/z 207[M+H]⁺

Synthesis of 360

A mixture of 359 (2.54 g, 12.29 mmol), 315 (4.36 g, 15.6 mmol),potassium carbonate (5.10 g, 36.9 mmol), tetrakistriphenylphosphinepalladium (1.42 g, 1.23 mmol) and water (2.2 ml)-1,2-dimethoxyethane(100 ml) was stirred under an argon atmosphere at 80° C. for 16 hours.The reaction solution was allowed to return to room temperature, dilutedwith ethyl acetate, dried over sodium sulfate and then filtered withcelite. The filtrate was concentrated to about 100 ml under reducedpressure, purified by silica gel column chromatography (eluting solvent:ethyl acetate/n-hexane=1/2) and then washed with ethyl acetate/n-hexane(=1/2) to obtain 360 (3.66 g, 92%) as an orange solid.

mp 175-175.5° C.

APCI-MS m/z 324[M+H]⁺

Synthesis of 361

A mixture of 360 (3.11 g, 9.62 mmol), lithium chloride (4.08 g, 96.2mmol) and hexamethylphosphoric tridamide (31 ml) was stirred under anargon atmosphere at 110° C. for 16 hours. The reaction solution wasallowed to return to room temperature, and ethyl acetate and water wereadded, and the solution was extracted with ethyl acetate. The extractionliquid was washed with water and dried, and then the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/2)and then recrystallized from ethyl acetate/n-hexane to obtain 361 (2.97g, 100%) as brown crystals.

mp 224-225° C.

APCI-MS m/z 310[M+H]⁺

Synthesis of 363

To a mixture of 361 (1.00 g, 3.23 mmol), 362 (1.40 g, 3.88 mmol),triphenylphosphine (1.02 g, 3.88 mmol) and tetrahydrofuran (40 ml), atetrahydrofuran (10 ml) solution of diisopropyl azodicarboxylate (0.77ml, 3.88 mmol) was added dropwise over 20 minutes under ice cooling,followed by stirring at the same temperature for 1 hour and furtherstirring at room temperature for 16 hours. To the reaction solution, 362(700 mg, 1.94 mmol), triphenylphosphine (510 mg, 1.94 mmol), diisopropylazodicarboxylate (0.38 ml, 1.92 mmol) and tetrahydrofuran (10 ml) wereadded, and the solution was further stirred at room temperature for 3hours. The solvent of the reaction solution was distilled off underreduced pressure, and the residue was purified by silica gel columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/9, 1/4) andthen recrystallized from ethyl acetate/n-hexane=1/4 to obtain 363 (1.94g, 92%) as orange crystals.

mp 144-145° C.

APCI-MS m/z 652 [M+H]+

Synthesis of 364

A mixture of 363 (1.00 g, 1.53 mmol), 10% Pd—C (moisture of about 50%;130 mg), ammonium formate (965 mg, 15.3 mmol) and methanol (20ml)-tetrahydrofuran (10 ml) was stirred under an argon atmosphere,followed by stirring at room temperature for 27 hours. The catalyst wasremoved by filtration and the solvent of the filtrate was distilled offunder reduced pressure. The residue was purified by silica gel columnchromatography (eluting solvent: n-hexane/ethyl acetate=4/1, 2/1) toobtain 364 (900 mg, 94%) as an orange amorphous.

APCI-MS m/z 622[M+H]⁺

Synthesis of 365

364 (890 mg, 1.43 mmol) and an aqueous 35% formaldehyde solution (0.61ml, 7.16 mmol) were nearly dissolved in methanol (50 ml)-tetrahydrofuran(10 ml) and magnesium sulfate (10 g) was added, and the mixture wasstirred at room temperature for 1 hour. The reaction solution wasice-cooled and NaBH₄ (271 mg, 7.16 mmol) was added at the sametemperature for 10 minutes, followed by stirring at room temperature for16 hours. Insolubles were removed from the reaction solution byfiltration, and the solvent of the filtrate was distilled off underreduced pressure. The residue was nearly dissolved in isopropyl alcohol(50 ml), and an aqueous 35% formaldehyde solution (0.61 ml, 7.16 mmol)and magnesium sulfate (10 g) were added, followed by stirring at roomtemperature for 30 minutes. The reaction solution was ice-cooled andNaBH₄ (271 mg, 7.16 mmol) was added. After stirring at room temperaturefor 2 days, NaBH₄ (271 mg, 7.16 mmol) was added, followed by furtherstirring at room temperature for 6 days. Insolubles were removed fromthe reaction solution by filtration and the solvent of the filtrate wasdistilled off under reduced pressure. The residue was dissolved in ethylacetate, and the solution was washed with water and dried, and then thesolvent was distilled off under reduced pressure. The residue waspurified by silica gel column chromatography (eluting solvent: ethylacetate/n-hexane=1/4) to obtain 365 (650 mg, 71%) as a pale yellowamorphous.

APCI-MS m/z 636[M+H]⁺

Synthesis of 366

To chloroform (12 ml) solution of 365 (640 mg, 1.01 mmol),trifluoroacetic acid (8 ml) was added dropwise under ice cooling andstirring, and water (2 ml) was added, and then the mixture was stirredat room temperature for 2 days. To the reaction solution, ice water andthen ethyl acetate were added, and the solution was extracted with ethylacetate after adjusting the pH to 9 using an aqueous potassium carbonatesolution. The extraction liquid was dried and the solvent was distilledoff under reduced pressure and the residue was purified by silica gelcolumn chromatography (eluting solvent: ethyl acetate/n-hexane=1/1) toobtain 366 (520 mg, 99%) as an orange amorphous.

APCI-MS m/z 522[M+H]⁺

Synthesis of THK-5163

To a methylene chloride (20 ml) solution of 366 (510 mg, 0.98 mmol) and3,4-dihydro-2H-pyran (1.77 ml, 19.6 mmol), paratoluenesulfonic acidmonohydrate (387 mg, 2.25 mmol) was added, and the mixture was stirredat room temperature for 20 minutes. After adjusting the pH of thereaction solution to 8 by adding triethylamine, the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel column chromatography (eluting solvent: ethyl acetate/n-hexane=1/2,2/3, 1/1) to obtain THK-5163 (569 mg, 96%) as an orange amorphous.

¹H NMR (400 MHz, DMSO-d₆) δ 1.32-1.51 (4H, m), 1.52-1.74 (2H, m), 2.34(3H, s), 2.78 (3H, d, J=3.6 Hz), 3.34-3.46 (1H, m), 3.64-3.72, 3.78-3.86(1H, m), 4.10 (2H, d, J=5.4 Hz), 4.14-4.22 (1H, m), 4.29-4.44 (2H, m),4.65-4.70, 4.85-4.88 (1H, m), 5.31 (1H, br), 6.57 (1H, dd, J=8.3, 1.4Hz), 6.93 (1H, d, J=2.1 Hz), 7.39 (2H, d, J=7.9 Hz), 7.44 (1H, dd,J=9.1, 2.4 Hz), 7.61-7.64 (1H, m), 7.70 (1H, d, J=8.2 Hz), 7.79 (2H, dd,J=8.3, 2.9 Hz), 7.83 (1H, d, J=9.4 Hz), 7.92 (1H, d, J=8.8 Hz), 8.08(1H, d, J=8.5 Hz)

IR (Nujol) 3433, 1733, 1619 cm⁻¹

APCI-MS m/z 606 [M+H]+

Synthesis Method of THK-5164

Synthesis of 367

To a mixture of 361 (1.09 g, 3.52 mmol), 311 (1.04 g, 5.0 mmol),triphenylphosphine (1.57 g, 5.98 mmol) and tetrahydrofuran (25 ml), atetrahydrofuran (5 ml) solution of diisopropyl azodicarboxylate (1.21 g,5.98 mmol) was added dropwise under ice cooling and stirring, followedby stirring at the same temperature for 1 hour and further stirring atroom temperature for 16 hours. The reaction solution was purified by NHsilica gel flash column chromatography (eluting solvent: ethylacetate/n-hexane=1/9) to obtain 367 (1.66 g, 94%) as an orange solid.

mp 119-120° C.

APCI-MS m/z 500[M+H]⁺

Synthesis of 368

A mixture of 367 (1.65 g, 3.3 mmol), 10% Pd—C (moisture of about 50%;350 mg), ammonium formate (2.08 g, 33 mmol) and methanol (40ml)-tetrahydrofuran (20 ml) was stirred under an argon atmosphere atroom temperature for 2 hours. The catalyst was removed by filtration andthe solvent of the filtrate was distilled off under reduced pressure.The residue was dissolved in ethyl acetate-tetrahydrofuran, and thesolution was washed in turn with water and saturated saline and dried,and then the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel flash column chromatography (elutingsolvent: n-hexane/ethyl acetate=3/1) to obtain 368 (1.40 g, 90%) as apale yellow solid.

mp 90-92° C.

APCI-MS m/z 470[M+H]⁺

Synthesis of 369

To a mixture of 368 (800 mg, 1.7 mmol), an aqueous 36% formaldehydesolution (708 mg, 8.5 mmol) and isopropanol (20 ml)-tetrahydrofuran (5ml), magnesium sulfate (5 g) was added, followed by stirring at roomtemperature for 1 hour. To the reaction solution, NaBH₄ (323 mg, 8.5mmol) was added, and the solution was stirred at room temperature for 24hours. Isopropanol (5 ml) and NaBH₄ (323 mg, 8.5 mmol) were added,followed by stirring at 80° C. for 1 hour. The reaction solution wasallowed to return to room temperature and insolubles were removed byfiltration, and the filtrate was purified by silica gel flash columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/5) to obtain369 (720 mg, 88%) as a pale yellow solid.

mp 134-135° C.

APCI-MS m/z 484[M+H]⁺

Synthesis of THK-5164

A mixture of 369 (700 mg, 1.45 mmol) and 48% HBr (7 ml) was stirred atroom temperature for 30 minutes. The reaction solution was poured intoice water, and the solution was made basic with concentrated ammoniawater and then extracted with ethyl acetate. The extraction liquid waswashed with saturated saline and dried, and then the solvent wasdistilled off under reduced pressure. The residue was washed with ethylacetate/n-hexane=1/5 and then recrystallized from ethyl acetate toobtain THK-5164 (490 mg, 91%) as a pale yellow solid.

mp 193-194° C.

¹H NMR (400 MHz, CDCl₃) δ 2.75 (1H, br), 2.92 (3H, s), 3.08 (6H, s),4.25-4.38 (3H, m), 4.40 (1H, br), 4.52-4.62 (1H, m), 4.65-4.74 (1H, m),6.69 (1H, d, J=8.2 Hz), 6.81 (1H, d, J=2.7 Hz), 7.35 (1H, dd, J=9.4, 2.7Hz), 7.63 (1H, dd, J=8.5, 1.8 Hz), 7.70 (1H, d, J=8.8 Hz), 7.79 (1H, d,J=1.8 Hz), 7.96 (1H, d, J=8.8 Hz), 8.00 (1H, brd, J=9.4 Hz)

IR (Nujol) 3417, 3250, 1620, 1592 cm⁻¹

APCI-MS m/z 370[M+H]⁺

Synthesis Method of THK-5165

Synthesis of 370

To an isopropanol (50 ml) solution of 355 (836 mg, 1.78 mmol), anaqueous 35% formaldehyde solution (760 mg, 8.9 mmol) and magnesiumsulfate (10 g) were added, and the mixture was stirred at roomtemperature for 2 hours. To the reaction solution, NaBH₄ (340 mg, 9mmol) was added three times every 24 hours, and the solution was stirredat room temperature for 2 days and then heated at reflux for 1 hour. Thereaction solution was allowed to return to room temperature andinsolubles were removed by filtration, and then the solvent of thefiltrate was distilled off under reduced pressure. The residue waspurified by silica gel flash column chromatography (eluting solvent:ethyl acetate/n-hexane=1/2, 2/5) to obtain 370 (683 mg, 84%) as a paleyellow amorphous.

APCI-MS m/z 456[M+H]⁺

Synthesis of THK-5165

A mixture of 370 (676 mg, 1.48 mmol) and 1M tetra-n-butylammoniumfluoride/tetrahydrofuran (5.0 ml, 5.0 mmol) was stirred at roomtemperature for 1 hour. The solvent of the reaction solution wasdistilled off under reduced pressure, and the residue was purified bysilica gel flash column chromatography (eluting solvent:chloroform→chloroform/methanol=50/1, 24/1) and then washed withn-hexane-diisopropyl ether to obtain THK-5165 (453 mg, 85%) as an orangesolid.

mp 79-82° C.

¹H NMR (400 MHz, DMSO-d₆) δ 2.80 (3H, d, J=4.9 Hz), 2.80 (3H, s),4.00-4.20 (3H, m), 4.50-4.70 (2H, m), 5.45-5.55 (4H, m), 6.56 (1H, d,J=8.2 Hz), 6.77 (1H, d, J=2.5 Hz), 7.11 (1H, dd, J=8.8, 2.5 Hz),7.60-7.70 (3H, m), 7.80 (1H, d, J=8.8 Hz), 7.90 (1H, d, J=8.8 Hz)

IR (Nujol) 1503, 1462, 1377 cm⁻¹

APCI-MS m/z 342[M+H]⁺

Synthesis Method of THK-5154

Synthesis of 347

A mixture of 346 (1.40 g, 6 mol), 314 (1.53 g, 6 mol), potassium acetate(2.36 g, 24 mol), PdCl₂(dppf) CH₂Cl₂ (245 mg, 0.3 mmol) and 1,4-dioxane(25 ml) was stirred under an argon atmosphere at 100° C. for 16 hours.The reaction solution was allowed to return to room temperature andinsolubles were removed by filtration, and the solvent of the filtratewas distilled off under reduced pressure. The residue was purified bysilica gel column chromatography (eluting solvent: n-hexane/ethylacetate=2/1) and then washed with n-hexane to obtain 347 (1.17 g, 70%)as a colorless solid.

mp 130-131° C.

APCI-MS m/z 281 [M+H]⁺

Synthesis of 348

A mixture of 347 (1.10 g, 3.39 mmol), 329 (840 mg, 4 mmol), sodiumcarbonate (850 mg, 8 mmol), tetrakistriphenylphosphine palladium (230mg, 0.2 mmol) water (2 ml) and 1,2-dimethoxyethane (20 ml) was stirredunder an argon atmosphere at 90° C. for 4 hours. The reaction solutionwas allowed to return to room temperature, and insolubles were collectedby filtration and then dried to obtain 348 (836 mg, 66%).

mp 253-255° C.

APCI-MS m/z 327 [M+H]⁺

Synthesis of 349

A mixture of 348 (326 mg, 1 mmol), lithium chloride (424 mg, 10 mmol)and hexamethylphosphoric triamide (7 ml) was stirred under an argonatmosphere at 110° C. for 19 hours. The reaction solution was allowed toreturn to room temperature and water was added, and the solution wasacidified with an aqueous citric acid solution and then extracted withethyl acetate. The extraction liquid was washed in turn with water andsaturated saline, and dried, and then the solvent was distilled offunder reduced pressure. The residue was washed with diisopropyl ether toobtain 349 (312 mg, 100%).

APCI-MS m/z 313[M+H]⁺

Synthesis of 350

To a mixture of 349 (680 mg, 2.18 mmol), 311 (680 mg, 3.26 mmol),triphenylphosphine (973 mg, 3.71 mmol) and tetrahydrofuran (20 ml), atetrahydrofuran (2 ml) solution of diisopropyl azodicarboxylate (750 mg,3.71 mmol) was added dropwise under ice cooling and stirring, followedby stirring at room temperature for 20 hours. The reaction solution waspurified by silica gel flash column chromatography (eluting solvent:ethyl acetate/n-hexane=1/9-1/5) and then washed with diisopropyl etherto obtain 350 (820 mg, 75%) as a grayish white solid.

mp 167-168° C.

APCI-MS m/z 503[M+H]⁺

Synthesis of 351

A mixture of 350 (510 mg, 1.01 mmol), Fe (503 mg), NH₄Cl (325 mg, 6.09mmol) and water (3 ml)-ethanol (15 ml) was stirred at 70 to 75° C. for1.5 hours. The reaction solution was allowed to return to roomtemperature, and then diluted with ethyl acetate. Insolubles wereremoved by filtration with celite and the filtrate was washed in turnwith ammonia water and saturated saline, and dried, and then the solventwas distilled off under reduced pressure. The residue was purified bysilica gel flash column chromatography (eluting solvent: chloroform,chloroform/methanol=40/1) to obtain 351 (385 mg, 86%) as an orangesolid.

mp 139-141° C.

APCI-MS m/z 443[M+H]⁺

Synthesis of 352

To a mixture of 351 (520 mg, 1.17 mmol), an aqueous 36% formaldehydesolution (1.95 g, 23.4 mmol) and ethanol (30 ml)-acetic acid (3 ml), apicoline borane complex (751 mg, 7.02 mmol) was added little by little,followed by stirring at room temperature for 4 hours. The reaction wascompleted by adding an aqueous 36% formaldehyde solution (3 g, 27.8mmol), acetic acid (1 ml) and a picoline borane complex (1.05 g, 9.82mmol). The reaction solution was made basic with concentrated ammoniawater under ice cooling and then extracted with ethyl acetate. Theextraction liquid was washed in turn with water and saturated saline,and dried, and then the solvent was distilled off under reducedpressure. The residue was purified by silica gel flash columnchromatography (eluting solvent: ethyl acetate/n-hexane=1/5) to obtain352 (430 mg, 74%) as a yellow solid.

APCI-MS m/z 499[M+H]⁺

mp 136-137° C.

Synthesis of THK-5154

A mixture of 352 (300 mg, 0.68 mmol) and 48% HBr (3 ml) was stirred atroom temperature for 10 minutes. The reaction solution was diluted withwater, and the solution was made basic with concentrated ammonia waterand then extracted with ethyl acetate. The extraction liquid was washedin turn with water and saturated saline, and dried, and then the solventwas distilled off under reduced pressure. The residue was washed withethyl acetate to obtain THK-5154 (220 mg, 84%) as a yellow solid.

mp 154-155° C.

¹H NMR (400 MHz, CDCl₃) δ 3.06 (6H, s), 3.10 (6H, s), 3.26 (1H, brs),4.24-4.32 (3H, m), 4.51-4.60 (1H, m), 4.64-4.72 (1H, m), 6.81 (1H, d,J=2.7 Hz), 7.37 (1H, dd, J=9.3, 2.7 Hz), 7.70 (1H, d, J=8.8 Hz), 7.98(1H, d, J=8.8 Hz), 7.98 (1H, d, J=9.3 Hz), 8.04 (1H, d, J=1.9 Hz), 8.53(1H, d, J=1.9 Hz).

IR (Nujol) 1618, 1588 cm⁻¹

APCI-MS m/z 385[M+H]⁺

Synthesis Method of THK-5166

Synthesis of 373

A mixture of 371 (654 mg, 4 mmol), 372 (1.28 g, 4.8 mmol), 2M sodiumcarbonate (4 ml, 8 mmol), tetrakistriphenylphosphine palladium (231 mg,0.2 mmol) and 1,2-dimethoxyethane (7 ml) was stirred under an argonatmosphere at 90° C. for 16 hours. The reaction solution was allowed toreturn to room temperature and the solvent was distilled off. Chloroformwas added and insolubles were removed by filtration. The chloroformlayer of the filtrate was separated and dried, and then the solvent wasdistilled off under reduced pressure. The residue was purified by silicagel flash column chromatography (eluting solvent: n-hexane,chloroform/n-hexane=1/4, 2/3, 3/2) and then washed with n-hexane toobtain 373 (986 mg, 92%) as a pale yellow solid.

APCI-MS m/z 269[M+H]⁺

Synthesis of 374

To a N,N-dimethylformamide (15 ml) solution of 373 (982 mg, 3.6 mmol)and 2-methanesulfonylethanol (679 mg, 5.47 mmol), 60% NaH (438 mg, 10.9mmol) was added at room temperature under stirring, and the mixture wasstirred at room temperature for 2 hours. After the reaction solution wasacidified by adding water and 10% hydrochloric acid water, the solutionwas made basic with an aqueous saturated sodium hydrogen carbonatesolution and then extracted with ethyl acetate. The extraction liquidwas washed with saturated saline and dried, and then the solvent wasdistilled off under reduced pressure. The residue was recrystallizedfrom ethyl acetate/n-hexane to obtain 374 (778 mg, 80%) as pale browncrystals.

APCI-MS m/z 267[M+H]⁺

Synthesis of 375

To a mixture of 374 (773 mg, 2.9 mmol), 311 (844 mg, 4.0 mmol),triphenylphosphine (1.21 g, 4.6 mmol) and tetrahydrofuran (10 ml), atetrahydrofuran (5 ml) solution of diisopropyl azodicarboxylate (936 mg,4.6 mmol) was added dropwise under ice cooling and stirring, followed bystirring at room temperature for 16 hours. The solvent of the reactionsolution was distilled off under reduced pressure, and the residue waspurified by silica gel flash column chromatography (eluting solvent:n-hexane, ethyl acetate/n-hexane=1/4) and then washed with n-hexane toobtain 375 (1.10 g, 83%) as a colorless solid.

APCI-MS m/z 457[M+H]⁺

Synthesis of 376

A mixture of 375 (1.09 g, 2.4 mmol), 10% Pd—C (moisture of about 50%;200 mg), ammonium formate (1.50 g, 24 mmol) and methanol (20ml)-tetrahydrofuran (10 ml) was stirred under an argon atmosphere atroom temperature for 1 hour. The catalyst was removed by filtration, andthe solvent of the filtrate was distilled off under reduced pressure. Tothe residue, water was added and the solution was extracted fromchloroform. The extraction liquid was dried and solvent was distilledoff under reduced pressure to obtain 376 (1.02 g, 100%) as a yellow oilysubstance.

APCI-MS m/z 427[M+H]⁺

Synthesis of 377

To a mixture of 376 (1.02 g, 2.4 mmol), an aqueous 35% formaldehydesolution (1.0 g, 11.7 mmol) and methanol (20 ml)-acetic acid (2 ml), apicoline borane complex (385 mg, 3.6 mmol) was added little by littleunder ice cooling and stirring, followed by stirring at room temperaturefor 2.5 hours. To the reaction solution, an aqueous 35% formaldehydesolution (0.5 g, 5.8 mmol) and a picoline borane complex (260 mg, 2.4mmol) were added, and the solution was further stirred at roomtemperature for 16 hours. The solvent was distilled off from thereaction solution under reduced pressure, and an aqueous saturatedsodium hydrogen carbonate solution was added to the residue, and thenthe solution was extracted with chloroform. The extraction liquid wasdried and the solvent was distilled off under reduced pressure. Theresidue was purified by silica gel flash column chromatography (elutingsolvent: n-hexane, ethyl acetate/n-hexane=1/20, 1/12) and then washedwith cold n-hexane to obtain 377 (827 mg, 74%) as a colorless solid.

APCI-MS m/z 455[M+H]⁺

Synthesis of THK-5166

A mixture of 377 (802 mg, 1.76 mmol) and 1M tetra-n-butylammoniumfluoride/tetrahydrofuran (5.0 ml, 5.0 mmol) was stirred at roomtemperature for 1 hour. The solvent of the reaction solution wasdistilled off under reduced pressure. The residue was purified by silicagel flash column chromatography (eluting solvent:chloroform→chloroform/methanol=50/1), purified by silica gel flashcolumn chromatography (eluting solvent: n-hexane→ethylacetate/n-hexane=1/1, 3/2) and then washed with n-hexane to obtain

THK-5166 (453 mg, 75%) as a pale yellow solid.

mp 81-84° C.

¹H NMR (400 MHz, DMSO-d₆) δ 2.86 (6H, s), 4.10-4.20 (3H, m), 4.40-4.70(2H, m), 5.54 (1H, brs), 7.04 (1H, brd, J=7.5 Hz), 7.54-7.58 (1H, tlike), 7.74-7.78 (1H,t like), 7.83 (1H, dd, J=8.5, 1.8 Hz), 7.87 (1H, d,J=1.8 Hz), 7.97 (1H, d, J=7.6 Hz), 8.05 (1H, d, J=8.5 Hz), 8.14 (1H, d,J=8.8 Hz), 8.40 (1H, d, J=8.8 Hz)

IR (Nujol) 1595, 1497, 1457, 1436 cm⁻¹

APCI-MS m/z 341[M+H]⁺

Synthesis Method of THK-5167

Synthesis of 378

To a mixture of 310 (452 mg, 1.54 mmol), 362 (560 mg, 1.55 mmol),triphenylphosphine (630 mg, 2.4 mmol) and tetrahydrofuran (20 ml), atetrahydrofuran (2 ml) solution of diisopropyl azodicarboxylate (485 mg,2.4 mmol) was added dropwise under ice cooling and stirring, followed bystirring at the same temperature for 20 minutes and further stirred atroom temperature for 16 hours. To the reaction solution, 362 (250 mg,0.69 mmol), triphenylphosphine (320 mg, 1.27 mmol) and diisopropylazodicarboxylate (250 mg, 1.24 mmol) were added, and the solution wasfurther stirred at room temperature for 20 hours. The reaction solutionwas purified by NH silica gel flash column chromatography (elutingsolvent: ethyl acetate/n-hexane=1/4, 1/3, 1/2, ethyl acetate,chloroform/methanol=10/1) and then washed with n-hexane/ethyl acetate toobtain 378 (683 mg, 70%).

mp 135-138° C.

Synthesis of 379

A mixture of 378 (700 mg, 1.1 mmol), tetrahydrofuran (15 ml), water (5ml) and trifluoroacetic acid (10 ml) was stirred at room temperature for16 hours. The reaction solution was diluted with ethyl acetate, and thesolution was washed in turn with an aqueous saturated sodium hydrogencarbonate solution and saturated saline, and dried and then the solventwas distilled off under reduced pressure. The residue was purified bysilica gel flash column chromatography (eluting solvent: ethylacetate/n-hexane=1/2, 1/1, 2/1) to obtain 379 (490 mg, 85%) as a solid.

mp 156-158° C.

APCI-MS m/z 522[M+H]⁺

Synthesis of THK-5167

To a methylene chloride (30 ml)-tetrahydrofuran (20 ml) solution of 379(480 mg, 0.92 mmol) and 3,4-dihydro-2H-pyran (1.55 g, 18.4 mmol),paratoluenesulfonic acid monohydrate (200 mg, 1.16 mmol) was added wasadded, and the mixture was stirred at room temperature for 78 hours. Thereaction solution was diluted with ethyl acetate (200 ml), and thesolution was washed in turn with an aqueous saturated sodium hydrogencarbonate solution and saturated saline, and dried, and then the solventwas distilled off under reduced pressure. The residue was purified bysilica gel flash column chromatography (eluting solvent: ethylacetate/n-hexane=1/2, 2/3, 1/1) and then recrystallized from ethylacetate to obtain THK-5167 (380 mg, 68%) as a pale yellow solid.

mp 153-155° C.

¹H NMR (400 MHz, CDCl₃) δ 1.45-1.62 (4H, m), 1.66-1.84 (2H, m), 2.34,2.35 (3H, each s), 3.21 (3H, s), 3.44-3.55 (3H, m), 3.83, 3.92 (1H, eachm), 4.06-4.39 (7H, m), 4.74, 4.83 (1H, each brt), 6.98, 7.00 (1H, eachd, J=2.7 Hz), 7.18, 7.19 (1H, each dd, J=9.1, 2.5 Hz), 7.227 (1H, d,J=8.2 Hz), 7.234 (1H, d, J=8.2 Hz), 7.73, 7.74 (1H, each d, J=9.4 Hz),7.75 (1H, brd, J=8.3 Hz), 7.77 (1H, brd, J=8.3 Hz), 7.836, 7.838 (1H,each d, J=1.8 Hz), 7.95 (1H, d, J=9.4 Hz), 8.00, 8.01 (1H, each d, J=8.8Hz), 8.486, 8.489 (1H, each d, J=1.8 Hz)

IR (Nujol) 1621, 1598 cm⁻¹

APCI-MS m/z 606[M+H]⁺

Synthesis Methods of THK-5168, THK-5170, THK-5171, THK-5172, THK-5173,THK-5174, THK-5175, THK-5176, THK-5179, THK-5181 and THK-5182

Each synthesis method is the same as any one of the above-mentionedsynthesis methods.

EXAMPLE 2 Labeling Synthesis of [¹⁸F] THK-5035

¹⁸F⁻ was synthesized by irradiating [¹⁸O] H₂O having isotope purity of95 or more with 12 MeV of proton beam accelerated by Cyclotron HM12(manufactured by Sumitomo Heavy Industries, Ltd.). Subsequently, thesolution thereof was passed through an anion-exchange resin (AG1-X8)thereby trapping ¹⁸F⁻ on the resin, followed by elution with a 33 mMK₂CO₃ solution. After transferring this aqueous ¹⁸F⁻-containing K₂CO₃solution (200 μL, 3.5 GBq) in a brown vial, Kryptofix 222 (16 mg) andacetonitrile (1.5 mL) were added and a He gas was sprayed while heatingin an oil bath (110° C.), and then acetonitrile was completely removedwhile azeotropically distilling water. Furthermore, an operation ofadding acetonitrile (1 mL) and removing acetonitrile in the same mannerunder heating conditions was repeated, thereby turning the state insidethe vial into a substantially moisture-free state. A DMSO solution (0.7mL) containing THK-5039 (4 mg), as a label precursor, dissolved thereinwas added, followed by heating and stirring in the oil bath (110° C.)for 10 minutes. Thereafter, the reaction solution was diluted withdistilled water (7 mL) and loaded into a Sep-Pack tC18 cartridge(manufactured by Waters) and, after washing the cartridge with distilledwater, the crude product was eluted with ethanol. A portion of theethanol solution was diluted with distilled water and subjected tosemi-preparative high-performance liquid chromatography (column:YMC-Pack Pro C18 RS (10×250 mm), mobile phase: MeCN/20 mM NaH₂PO₄=60/40,flow rate: 5.0 mL/min) and then [¹⁸F] THK-5035-derived radioactive peak(121 MBq, no decay correction), which is eluted within about 14 to 15minutes, was dispensed. After the fraction was diluted with distilledwater, [¹⁸F] THK-5035 was subjected to solid-phase extraction using aSep-Pak tC18 cartridge and then eluted with ethanol. In anautoradiography test, the ethanol solution was used after appropriatelybeing diluted. In a biodistribution test, Polysorbate 80 was added tothe ethanol solution and, after distilling off ethanol by an evaporatorethanol, a radioactive residue containing [¹⁸F] THK-5035 in the flaskwas dissolved in physiological saline and then used as an injection in abrain migration evaluation test.

Staining Test on Compounds of the Present Invention on Brain Section ofAlzheimer's Disease Patients

The procedure of staining test on the compounds of the present inventionon the brain section of Alzheimer's disease patients is described below:

(1) We used the brain specimens of the temporal lobe or hippocampus frompatients who were definitely diagnosed as having Alzheimer's diseasepathology. The specimens were obtained from Choju Medical Institute,Fukushimura Hospital, our collaborating research institute, and theconsent for use for research purposes was obtained from the patients'bereaved family (Fukushimura Hospital, Ethics Committee Approval No.20).

(2) The paraffin-embedded brain tissue was sliced to 6 or 8 μmthickness, extended on a slide glass, and dried. The paraffinized brainsection was deparaffinized by washing with xylene for 10 minutes×2, 100%ethanol for 5 minutes×2, 90% ethanol for 5 minutes, and running waterfor 10 minutes in this order.

(3) As a pretreatment for staining with the compounds of the presentinvention, the treatment to remove autofluorescence with lipofuscin wasperformed. First, the paraffinized brain section was immersed in 0.25%KM_(n)O₄ solution for 20 minutes. It was washed with PBS for 2 minutestwice, immersed in a 0.1% K₂S₂O₅/oxalic acid solution for approximately5 minutes, and further washed with PBS for 2 minutes 3 times.

(4) From 100 μM solution of the compounds of the present inventiondissolved in 50% ethanol, approximately 150 μl was dropped on thesection to react for 10 minutes. After being immersed in tap water 5times, it was mounted with Flour Save Reagent (Calbiochem) and examinedwith a fluorescence microscope (Nikon, ECLIPSE 80i). The images weretaken with a digital camera (Nikon, Dxin1200F or Photometrics, Cool SNAPES).

The results of above mentioned staining test on the compounds of thepresent invention are illustrated in FIGS. 5 to 20. It was found thatall of THK-5035, THK-5038, THK-5058, THK-5064, THK-5065, THK-5066,THK-5071, THK-5077, THK-5078, THK-5079, THK-5080, THK-5081, THK-5082,THK-5087, THK-5088, THK-5089, THK-5091, THK-5092, THK-5097, THK-5098,THK-5059, THK-5075, THK-5076, THK-5086, THK-5100, THK-5105, THK-5106,THK-5107, THK-5112, THK-5116, THK-5117, and THK-932 bind toneurofibrillary tangles specifically and selectively in the brainsection of Alzheimer's disease patients, and that the compound ofFormula (I) of the present invention has high specificity to tau (FIGS.5 to 20).

The results of above mentioned staining test on the compounds of thepresent invention are illustrated in FIGS. 25 to 29. It was found thatall of THK-5136, THK-5153, THK-5157, THK-5128, THK-5147, THK-5155,THK-5156, THK-5164, and THK-5154 bind to neurofibrillary tanglesspecifically and selectively in the brain section of Alzheimer's diseasepatients, and that the compound of Formula (I) of the present inventionhas high specificity to tau (FIGS. 25 to 29).

Autoradiography Test

After the paraffin-embedded brain section of Alzheimer's diseasepatients was deparaffinized, it was immersed in PBS for 10 minutes.Approximately 400 μCi/ml of [¹⁸F] BF-227 and [¹⁸F] THK-5035 were droppedon the section to react for 10 minutes at room temperature. It wasimmersed in distilled water for 2 minutes, shaken in 50% EtOH lightlyfor 2 minutes, immersed again in distilled water for 2 minutes, anddried in a paraffin extension device. The section was contacted on animaging plate and settled overnight, and the next day, the images wereread with a BAS5000 (Fujifilm Corporation). Furthermore, the serialsection was immunostained with thioflavine-S staining and anti-tauantibody (AT8).

FIG. 21 illustrates the autoradiography images of [¹⁸F] BF-227 and [¹⁸F]THK-5035, the thioflavine-S (TF-S) staining images in the serialsection, and the anti-tau antibody (Tau) staining. In theautoradiography images, the region where [¹⁸F] THK-5035 was accumulated(region enclosed with rectangles) showed no accumulation of [¹⁸F] BF-227showing high affinity to amyloid β. In the region, abundant anti-tauantibody immunostaining positive structures were also observed. From themorphological images in the thioflavine-S staining images, which stainneurofibrillary tangles, it is strongly suggested that the structuresobserved in the region are neurofibrillary tangles. From the aboveresults, it was confirmed that [¹⁸F] THK-5035 shows higher affinity toneurofibrillary tangles (tau protein) than to amyloid β.

Evaluation of Mouse Brain Transition of Labeled Compounds

The physiologic saline containing [¹⁸F] THK-5035 was administered to thecaudate vein of male ICR mice (6-7 week old). By considering theaccumulation of radioactivity in the brain tissue and plasma tissue 2minutes after administration, we evaluated the brain uptake of labeledcompounds.

In the evaluation of radioactive accumulation, the ratio ofradioactivity per unit weight of the tissue to be evaluated to alladministered radioactivity (% Injected Dose/g of tissue; % ID/g) wasused as index. For measurement of radioactivity, a gamma counter (1480WIZARD, PerkinElmer, Inc.) was used. In the test, the labeled compoundswere administered to the caudate vein, and 2 minutes later, cervicaldislocation was performed to the mice under etherization. Then,immediately blood was collected from the heart, and the whole brain(including the cerebellum and brainstem) was extirpated. After then, theradioactivity and tissue weight of each sample were measured, and thedata was used to calculate the % ID/g.

Table 3 illustrates the results of this evaluation test.

TABLE 3 % ID/g or ml after 2 minutes (mean ± SD) [¹⁸F] THK-5035 Brain6.01 ± 0.54 Plasma 2.20 ± 0.26

The value of 0.5% ID/g or more is considered to be enough to indicatethe brain transition of labeled compounds for PET or SPECT targeting tothe central nervous system. From that viewpoint, it was found that the[¹⁸F] labeled compounds of the formula (I) of the present invention are[¹⁸F] labeled compounds having extremely high brain uptake.

Synthesis of Label, [¹⁸F] THK-5105

[¹⁸O]-H₂O with an isotope purity of 98% or higher was irradiated with a12 MeV proton beam, which was accelerated by a cyclotron HM 12 (SumitomoHeavy Industries, Ltd.), to produce ¹⁸F⁻. Subsequently, the resultingsolution was passed through an anion-exchange resin (AG 1-X8) to capturethe ¹⁸F⁻ on the resin, which was eluted with a 33 mM solution of K₂CO₃.Into an amber vial was placed 200 μL of the ¹⁸F⁻-containing aqueousK₂CO₃ solution (3.13 GBq), and Kryptofix 222 (16 mg) and acetonitrile(2.3 mL) were added. He gas was sprayed while the vial was heated in anoil bath (110° C.), so as to remove completely the acetonitrile withazeotropic evaporation of water. Another acetonitrile (1.5 mL) wasadded, and the acetonitrile was removed under heating, in a similar way.These procedures were repeated twice to make the inside of the vialanhydrous. To the vial was added a DMSO solution (0.70 mL) of a labelprecursor THK-5121 (3.0 mg), and the mixture was heated and stirred for10 minutes in an oil bath (110° C.). After that, hydrochloric acid (2M,0.2 mL) was added to the reaction solution and an additional reactionwas carried out at 110° C. for 3 minutes. Then, the reaction solutionwas diluted with a potassium acetate solution (4M, 0.1 mL) and distilledwater (7.0 mL), and loaded on a Sep-Pak tC18 cartridge (Waters), whichwas then washed with distilled water, followed by elution of crudeproduct with ethanol. An ethanol-eluted fraction with the highestradioactivity was diluted with distilled water and subjected tosemi-preparative high performance liquid chromatography using anInertsil ODS-4 column (10×250 mm), a mobile phase of MeCN/NaH₂PO₄ (20mM) (50/50), and a flow rate of 5.0 mL/min. A radioactive peak of [¹⁸F]THK-5105, which was eluted at about 18.5 minutes (988 MBq, decayuncorrected), was collected. Analysis of the collected sample revealed aradiochemical purity of 98% or more and a specific radioactivity of 137GBq/μmol (on analysis).

The preparative HPLC fraction of [¹⁸F] THK-5105, which was synthesizedaccording to the synthesis procedure described above, was diluted withdistilled water, and then subjected to solid-phase extraction using aSep-Pak tC18 cartridge, followed by elution with ethanol or DMSO andappropriate dilution, and used in binding tests and autoradiographyexperiments. For experiments on its in vivo distribution (evaluation ofits brain delivery), Polysorbate 80 was added to the ethanol-elutedfraction, from which the ethanol was removed using an evaporator. The[¹⁸F] THK-5105 containing radioactive residue within the flask wasdissolved in physiological saline and the solution prepared was used asa solution for injection.

Synthesis of Label, [¹⁸F] THK-5117

[¹⁸F] THK-5117 was synthesized according to the above-described methodfor synthesis of label, [¹⁸F] THK-5105. In the synthesis of [¹⁸F]THK-5117, the synthesis was carried out using 500 μL of a¹⁸F⁻-containing aqueous K₂CO₃ solution (3.10 GBq), 3.0 mg of a labelprecursor THK-5119, and MeCN/NaH₂PO₄ (20 mM) (45/55) as the mobile phasein the semi-preparative high performance liquid chromatography, and wasable to give 770 MBq of [¹⁸F] THK-5117 (decay uncorrected). As in [¹⁸F]THK-5105, a solution for injection was prepared and used in experimentsfor the evaluation of its brain delivery.

Synthesis of Label, [¹⁸F] THK-5125

[¹⁸F] THK-5125 was synthesized according to the above-described methodfor synthesis of label, [¹⁸F] THK-5105. In the synthesis of [¹⁸F]THK-5125, the synthesis was carried out using 200 μL of a¹⁸F⁻-containing aqueous K₂CO₃ solution (3.14 GBq), 3.3 mg of a labelprecursor THK-5131, and MeCN/NaH₂PO₄ (20 mM) (45/55) as the mobile phasein the semi-preparative high performance liquid chromatography, and wasable to give 1.36 GBq of [¹⁸F] THK-5125 (decay uncorrected). As in [¹⁸F]THK-5105, a solution for injection was prepared and used in experimentsfor the evaluation of its brain delivery.

Synthesis of Label, [¹⁸F] FDDNP

Into an amber vial was placed 300 μL of a ¹⁸F⁻-containing aqueous K₂CO₃solution (3.87 GBq), which was prepared in a similar way as in [¹⁸F]THK-5105, and Kryptofix 222 (16 mg) and acetonitrile (2.3 mL) wereadded. He gas was sprayed while the vial was heated in an oil bath (110°C.), so as to remove completely the acetonitrile with azeotropicevaporation of water. More acetonitrile was added, and the acetonitrilewas removed under heating, in a similar way. These procedures wererepeated three times to make the inside of the vial anhydrous. To thevial was added an acetonitrile solution (0.70 mL) of a label precursor(2.7 mg), which has a tosyl group as a leaving group attached at theposition of the fluorine in FDDNP (Jie Liu et al., Molecular Imaging andBiology (2007), vol. 9, pp. 6-16). The mixture was heated and stirredfor 10 minutes in an oil bath (110° C.), in an unsealed state and withaddition of 0.1 mL of acetonitrile every other minute. After that, thereaction solution was air cooled, diluted with distilled water (7.0 mL),and loaded on a Sep-Pak tC18 cartridge (Waters), which was then washedwith distilled water, followed by elution of the crude product withethanol. An ethanol-eluted fraction with the highest radioactivity wasdiluted with distilled water and subjected to semi-preparative highperformance liquid chromatography using an Inertsil ODS-4 column (10×250mm), a mobile phase of MeCN/NaH₂PO₄ (20 mM) (60/40), and a flow rate of5.0 mL/min. A radioactive peak of [¹⁸F] FDDNP, which was eluted at about19.5 minutes (814 MBq, decay uncorrected), was collected. As in [¹⁸F]THK-5105, a solution for injection was prepared and used in experimentsfor the evaluation of its brain delivery.

Evaluation of Brain Delivery Of [¹⁸F] THK-5105, [¹⁸F] THK-5117, and[¹⁸F] THK-5125 in Mice

A physiological saline solution containing [¹⁸F] THK-5105,[¹⁸F]THK-5117, or [¹⁸F] THK-5125 was administered into the tail vein ofmale ICR mice (6 to 7 weeks old), and brain delivery of these labelcompounds was assessed from the accumulation of radioactivity in brainand plasma tissues at two minutes post-injection.

As an index in the evaluation of radioactive accumulation, use was madeof the percentage of radioactivity per unit weight of brain relative tothe total radioactivity administered (% Injected Dose/g of tissue; %ID/g). For measurement of radioactivity, a gamma counter (1480 WIZARD,Perkin Elmer) was used. Experimental procedures were as follows. A labelcompound was administered into the tail vein of mice. Two minutes later,each mouse was subjected to cervical dislocation under ether anesthesia,and the whole brain, including the cerebellum and brain stem, wasisolated immediately. The radioactivity and tissue weight of the samplewere measured, and their data were used to calculate a % ID/g values forthe labeled compound.

Table 4 shows % ID/g values at 2 minutes and at 60 minutespost-administration, and values of the % ID/g values at 2 minutespost-administration divided by those at 60 minutes post-administration.Labeled probes which target an intended histopathology in the brain,need to be incorporated rapidly into the brain and to be rapidly washedout from histopathologies that are not their intended histopathology.[¹⁸F] THK-5105, [¹⁸F] THK-5117 and [¹⁸F] THK-5125 exhibited a sufficientintracerebral content at 2 minutes post-administration, and thequotients of their intracerebral contents at 2 minutespost-administration and 60 minutes post-administration were greater thanthose for [¹⁸F] BAY94-9172 and [¹⁸F] AV-45, which are known as amyloidimaging probes, and that for [¹⁸F] FDDNP. Form these results, it turnedout that [¹⁸F] THK-5105, [¹⁸F] THK-5117, and [¹⁸F] THK-5125 weresuperior in being delivered to and washed out from the brain. (The dataof [¹⁸F] BAY94-9172 was taken from Zhang et al., Nuclear Medicine andBiology, 32, 799-809, 2005, and the data of [¹⁸F] AV-45 from Choi etal., J Nucl Med, 50, 1887-1894, 2009.)

TABLE 4 2 minutes post- % ID/g administration/ 2 minutes post- 60minutes post- 60 minutes post- administration administrationadministration [¹⁸F]FDDNP 6.23 2.14 2.91 [¹⁸F]BAY94- 7.77 1.61 4.83 9172[¹⁸F]AV-45 7.33 1.88 3.90 [¹⁸F]THK-5105 9.20 1.00 9.20 [¹⁸F]THK-51176.06 0.26 23.3 [¹⁸F]THK-5125 7.82 0.61 12.8

Autoradiography Experiments Using [¹⁸F] THK-5105, [¹⁸F] THK-5117, and[¹⁸F] THK-5125

Paraffin-embedded sections of the brain from an Alzheimer's diseasepatient were deparaffinized, and then immersed in PBS for 10 minutes.Approximately 400 μCi/ml of each of [¹⁸F] THK-5105, [¹⁸F] THK-5117, and[¹⁸F] THK-5125 was added dropwise to deparaffinized sections and allowedto react for 10 minutes at room temperature. After that, the sectionswere immersed in distilled water for 2 minutes, shaken lightly in 50%EtOH for 2 minutes, immersed again in distilled water for 2 minutes, anddried in a paraffin stretching device. The section was left overnight incontact with an imaging plate. Next day, the image was read with a BAS5000 (Fujifilm Corporation). In addition, adjacent sections wereimmunostained with anti-phosphorylated tau antibody (AT8).

FIG. 22 shows an image of [¹⁸F] THK-5105 autoradiography of a section ofthe hippocampus and images of unlabeled THK-5105 staining and ofanti-phosphorylated tau antibody (AT8) staining of its adjacentsections. In the autoradiography image, areas where the accumulation of[¹⁸F] THK-5105 was observed were found abundant in structures which werepositive (for pTau) by immunostaining with the anti-phosphorylated tauantibody. Also in their respective higher magnification images, the[¹⁸F] THK-5105 autoradiography image corresponded well with the image ofimmunostaining with the anti-phosphorylated tau antibody, and further,the image of unlabeled THK-5105 staining corresponded well with theimage of immunostaining with the anti-phosphorylated tau antibody,including their morphological images.

FIG. 23 shows images of [¹⁸F] THK-5105, [¹⁸F] THK-5117, and [¹⁸F]THK-5125 autoradiography of sections of the lateral temporal cortex andof the medial temporal cortex. [¹⁸F] THK-5105, [¹⁸F] THK-5117, and [¹⁸F]THK-5125 were observed to be accumulated in the lateral temporal cortexand medial temporal cortex, which are known as areas where tauproduction occurs late in Alzheimer's disease.

From these results, it was ascertained that [¹⁸F] THK-5105, [¹⁸F]THK-5117, and [¹⁸F] THK-5125 have high affinity to neurofibrillarytangles (tau protein).

Evaluation of binding to Tau Protein Aggregates

Regarding the binding of tau imaging probes to tau protein aggregates, abinding test was carried out and evaluated using a K18-ΔK280 constructwhich contained a 4-repeat structure involved in the formation ofbeta-sheet structures in the htau40 protein. For K18-ΔK280 (Martin vonBergen et al., THE JOURNAL OF BIOLOGICAL CHEMISTRY (2001), vol. 276, pp.48165-48174; M. Goedert et al., NEURON (1989), vol. 3, pp. 519-526), anartificial gene fragment which was prepared based on its amino acidsequence was cloned into a pET-3a vector, the resulting plasmid was usedto transform Escherichia coli BL21 (DE3) competent cells for proteinexpression, and the recombinant Escherichia coli cell was cultured toexpress K18-ΔK280. The K18-ΔK280 expressed was purified according to themethod described in the literature (Stefan Barghorn et al., Methods inMolecular Biology, vol. 299, pp. 35-51). In the binding test, aggregatesof the purified K18-ΔK280 were made and used.

K18-ΔK280 aggregates were made by preparing 20 μM PBS (pH 7.4) andincubating the K18-ΔK280 in the PBS at 37° C. for 4 days. The statewhere beta-sheet structures in K18-ΔK280 aggregates were formed wasascertained by thioflavin S fluorescence binding test. The aggregatessolution was diluted to 400 nM in assay buffer (PBS, 0.1% BSA) and usedin the binding test.

Saturation Binding Test of THK-5105

In the binding test of [¹⁸F] THK-5105, reaction solutions were preparedsuch that K18-ΔK280 aggregates was at a final concentration of 200 nM inthe reaction system and [¹⁸F] THK-5105 was at final concentrations offrom 0.1 to 100 nM, and incubated at room temperature for one hour.After the incubation, [¹⁸F] THK-5105 binding to K18-ΔK280 aggregates andTHK-5105 not binding to K18-ΔK280 aggregates were separated using aMultiScreen HTS filter plate (96 wells, Millipore Corporation) and thefilter was washed with assay buffer (200 μL, 3 times). The total bindingof [¹⁸F] THK-5105 to K18-ΔK280 aggregates was calculated from theradioactivity of the filter which was measured on a gamma counter(AccuFLEX γ7000, Aloka). Non-specific binding was determined by addingunlabeled THK-5105 (at a final concentration of 2 μM) to the reactionsystem and performing experiments in a similar way. The data of thetotal binding and the non-specific binding were used and analyzed usingan analysis software GraphPad prism (Ver. 5), which gave a low value ofdissociation constant Kd of 3.9 nM, indicating that THK-5105 displayed ahigh binding affinity to K18-ΔK280 aggregates.

Competitive Binding Test Using [¹⁸F] THK-5105 as Radioactive Competitor

For tau imaging probes other than THK-5105, binding to K18-ΔK280aggregates was evaluated with a competitive binding test using [¹⁸F]THK-5105 as a radioactive competitor. In the binding test, [¹⁸F]THK-5105 (at a final concentration of 1.76 nM) and a tested compound (atfinal concentrations of from 0.1 to 1000 nM) coexisted in the reactionsystem, to which K18-ΔK280 aggregates (at a final concentration of 200nM) was then added and the mixtures were incubated at room temperaturefor one hour. The radioactivity of [¹⁸F] THK-5105 binding to theaggregates was determined in a similar way as in the saturation bindingtest of THK-5105. The percentage of [¹⁸F] THK-5105 bonding in theabsence of each tested compound was set to be 100%, and percentages of[¹⁸F] THK-5105 bonding at various concentrations of each of the testedcompounds were determined. From these data, the inhibition constants(Kis) of the respective tested compounds were calculated using ananalysis software GraphPad prism (Ver. 5). The results are shown inTable 5 and FIG. 24. Any of the tested compounds displayed a highbinding affinity to K18-ΔK280 aggregates.

TABLE 5 Ki (nM) THK-5116 36.0 THK-5117 10.5 THK-5122 34.9 THK-5125 9.2THK-5129 21.4 THK-5151 40.1 THK-523 (reference for comparison) 59.3FDDNP (reference for comparison) 256

INDUSTRIAL APPLICABILITY

The compounds of the present invention are very useful, for example, inearly detection, treatment and prevention of neurofibrillary tanglesincluding Alzheimer's disease, and can be utilized in the fields of theproduction of diagnostic agents and diagnostic kits for these diseases,the fields of the production of remedies and preventatives for thesediseases, studies of these diseases and the like.

1-43. (canceled)
 44. A compound of the formula (I):

wherein A is

R¹ is halogen, a —C(═O)-lower alkyl group (the alkyl group eachindependently may be substituted with one or more substituents selectedfrom NR^(a)R^(b), halogen and a hydroxy group), a lower alkyl group (thealkyl group each independently may be substituted with one or moresubstituents selected from halogen and a hydroxy group), a —O-loweralkyl group (the alkyl group each independently may be substituted withone or more substituents selected from halogen and a hydroxy group), or

in which R⁴ and R⁵ each independently represents hydrogen, a lower alkylgroup or a cycloalkyl group, or R⁴, R⁵ and the nitrogen atom to whichthey are attached are taken together to form a 3- to 8-memberednitrogen-containing aliphatic ring (one or more carbon atomsconstituting the nitrogen-containing aliphatic ring may be substitutedwith a nitrogen atom, a sulfur atom or an oxygen atom, and when a carbonatom is substituted with a nitrogen atom, the nitrogen atom may besubstituted with a lower alkyl group), or R⁴ and the nitrogen atom towhich it is attached are taken together with ring A to form a 8- to16-membered nitrogen-containing fused bicyclic ring (one or more carbonatoms constituting the nitrogen-containing fused bicyclic ring may besubstituted with a nitrogen atom, a sulfur atom or an oxygen atom, andwhen a carbon atom is substituted with a nitrogen atom, the nitrogenatom may be substituted with a lower alkyl group) and R⁵ representshydrogen, a lower alkyl group or a cycloalkyl group, in which the line,that the dotted line intersects, means a bond of the above generalformula to the other structural moiety, R² or R³ each independentlyrepresents halogen, OH, COOH, SO₃H, NO₂, SH, NR^(a)R^(b), a lower alkylgroup (the alkyl group each independently may be substituted with one ormore substituents selected from halogen and a hydroxy group) or a—O-lower alkyl group (the alkyl group each independently may besubstituted with one or more substituents selected from halogen and ahydroxy group), ring A is unsubstituted, or substituted with R⁶ (inwhich R⁶ is one or more substituents selected independently fromhalogen, OH, COOH, SO₃H, NO₂, SH, NR^(a)R^(b), a lower alkyl group (thealkyl group each independently may be substituted with one or moresubstituents selected from halogen and a hydroxy group) and a —O-loweralkyl group (the alkyl group each independently may be substituted withone or more substituents selected from halogen, a hydroxy group and a—O-lower alkyl group-O-lower alkyl group (the alkyl group eachindependently may be substituted with halogen)), R^(a) and R^(b) eachindependently represents hydrogen or a lower alkyl group (the alkylgroup each independently may be substituted with one or moresubstituents selected from halogen and a hydroxy group), m is an integerof 0 to 4, n is an integer of 0 to 4, wherein at least one of R², R³ andR⁶ is a —O-lower alkyl group substituted with one hydroxy group and onehalogen, or a pharmaceutically acceptable salt or solvate thereof. 45.The compound according to claim 44, wherein R¹ is halogen, a—C(═O)-lower alkyl group (the alkyl group each independently may besubstituted with NH₂), a lower alkyl group (the alkyl group eachindependently may be substituted with a hydroxy group), —O-lower alkylgroup, or

in which R⁴ and R⁵ each independently represents hydrogen or a loweralkyl group, or a pharmaceutically acceptable salt or solvate thereof.46. The compound according to claim 44, wherein at least one of R², R³and R⁶ is represented by:

or a pharmaceutically acceptable salt or solvate thereof.
 47. Thecompound according to claim 44, wherein at least one of R², R³ and R⁶ isNR^(a)R^(b), and R^(a) and R^(b) each independently represents hydrogenor an unsubstituted lower alkyl group, or a pharmaceutically acceptablesalt or solvate thereof.
 48. The compound according to claim 44, whereinthe compound is labeled, or a pharmaceutically acceptable salt orsolvate thereof.
 49. A pharmaceutical composition comprising thecompound according to claim 44, or a pharmaceutically acceptable salt orsolvate thereof.
 50. A composition for the diagnosis, treatment and/orprevention of conformational disease, comprising the compound accordingto claim 44, or a pharmaceutically acceptable salt or solvate thereof.51. A kit for the diagnosis of conformational disease, or detecting orstaining a beta-sheet structure protein comprising the compoundaccording to claim 44, or a pharmaceutically acceptable salt or solvatethereof as an essential ingredient.
 52. A method of treating and/orpreventing conformational disease, or diagnosing a conformationaldisease in a subject, which comprises administering the compoundaccording to claim 44, or a pharmaceutically acceptable salt or solvatethereof to the subject.
 53. A method of detecting or staining a β-sheetstructure protein in a sample, which comprises staining the sample usingthe compound according to claim 44, or a pharmaceutically acceptablesalt or solvate thereof.
 54. A method of producing the compound of theformula (I), which comprises the following steps of: (i) reacting acompound of the formula (II):

in which, R² and m are as defined in the formula (I), and R⁷ representsNH₂ or NO₂, with a compound of the formula (III):

in which A and R¹ are as defined in the formula (I), to obtain acompound of the formula (IV):

and isolating this compound as the compound of the formula (I), or (ii)optionally converting the compound of the formula (IV) into anothercompound of the formula (I) and isolating the compound.
 55. A method ofproducing the compound of the formula (I), which comprises the followingsteps of: (i) reacting a compound of the formula (V):

in which R², R³, m and n are as defined in the formula (I), and R⁸ is ahydroxyl group or halogen, provided that at least one of R² or R³ is ahydroxy group, with a compound of the formula: OH-Ark (Ark eachindependently represents a lower alkyl group which may be substitutedwith one or more substituents selected from the group consisting ofhalogen and a hydroxy group, to obtain a compound of the formula (V′):

in which R², R³, m and n are as defined in the formula (I), and R⁸ is ahydroxyl group or halogen, provided that at least one of R² or R³ is —O—Ark (Ark is as defined above), and (ii) reacting the compound of theformula (V′) with a compound of the formula (VI) or (VII):

in which A and R¹ are as defined in the formula (I), to obtain thecompound of the formula (I) in which at least one of R² and R³ is a—O-lower alkyl group (the alkyl group each independently may besubstituted with one or more substituents selected from the groupconsisting of halogen and a hydroxy group), or isolating the compound,or (iii) optionally converting the obtained compound of the formula (I)into another compound of the formula (I), and isolating the compound.56. A method of producing the compound of the formula (I) in which atleast one of R² and R³ is a —O-lower alkyl group (the alkyl group eachindependently may be substituted with one or more substituents selectedfrom the group consisting of halogen and a hydroxy group), whichcomprises the following steps of: (i) reacting a compound of the formula(V):

in which R², R³, m and n are as defined in the formula (I), and R⁸ is ahydroxyl group or halogen, provided that at least one of R² or R³ is ahydroxy group, with a compound of the formula (VI) or (VII):

in which A and R¹ are as defined in the formula (I), to obtain acompound of the formula (V″):

in which R¹, R², R³, A, m and n are as defined in the formula (I),provided that at least one of R² or R³ is a hydroxy group, and (ii)reacting the compound of the formula (V″) with a compound of theformula: OH-Ark (Ark each independently represents a lower alkyl groupwhich may be substituted with one or more substituents selected from thegroup consisting of halogen and a hydroxy group) to obtain a compound ofthe formula (I) in which at least one of R² and R³ is a —O-lower alkylgroup (the alkyl group each independently may be substituted with one ormore substituents selected from the group consisting of halogen and ahydroxy group), and isolating the compound, or (iii) converting theobtained compound of the formula (I) into another compound of theformula (I), and isolating the compound.
 57. A compound of the formula(I′):

wherein A is

R¹ is halogen, a —C(═O)-lower alkyl group (the alkyl group eachindependently may be substituted with one or more substituents selectedfrom NR^(a)R^(b), halogen and a hydroxy group), a lower alkyl group (thealkyl group each independently may be substituted with one or moresubstituents selected from halogen and a hydroxy group), a —O-loweralkyl group (the alkyl group each independently may be substituted withone or more substituents selected from halogen and a hydroxy group), or

in which R⁴ and R⁵ each independently represents hydrogen, a lower alkylgroup or a cycloalkyl group, or R⁴, R⁵ and the nitrogen atom to whichthey are attached are taken together to form a 3- to 8-memberednitrogen-containing aliphatic ring (one or more carbon atomsconstituting the nitrogen-containing aliphatic ring may be substitutedwith a nitrogen atom, a sulfur atom or an oxygen atom, and when a carbonatom is substituted with a nitrogen atom, the nitrogen atom may besubstituted with a lower alkyl group), or R⁴ and the nitrogen atom towhich it is attached are taken together with ring A to form a 8- to16-membered nitrogen-containing fused bicyclic ring (one or more carbonatoms constituting the nitrogen-containing fused bicyclic ring may besubstituted with a nitrogen atom, a sulfur atom or an oxygen atom, andwhen a carbon atom is substituted with a nitrogen atom, the nitrogenatom may be substituted with a lower alkyl group) and R⁵ representshydrogen, a lower alkyl group or a cycloalkyl group, in which the line,that the dotted line intersects, means a bond of the above generalformula to the other structural moiety, R² or R³ each independently maybe substituted with halogen, OH, COOH, SO₃H, NO₂, SH, NR^(a)R^(b), alower alkyl group (the lower alkyl group each independently may besubstituted with one or more substituents selected from ap-toluenesulfonyloxy group, a methanesulfonyloxy group, atrifluoromethanesulfonyloxy group or a 2-tetrahydropyranyloxy, halogenand a hydroxy group) or a —O-lower alkyl group (the lower alkyl group issubstituted with a p-toluenesulfonyloxy group, a methanesulfonyloxygroup, trifluoromethanesulfonyloxy group or a 2-tetrahydropyranyloxy,and also may be substituted with a hydroxy group), ring A isunsubstituted or substituted with R⁶ (in which R⁶ is one or moresubstituents selected independently from halogen, OH, COOH, SO₃H, NO₂,SH, NR^(a)R^(b), a lower alkyl group (the lower alkyl group eachindependently may be substituted with one or more substituents selectedfrom a p-toluenesulfonyloxy group, a methanesulfonyloxy group, atrifluoromethanesulfonyloxy group or a 2-tetrahydropyranyloxy, halogenand a hydroxy group) and an —O-lower alkyl group (the lower alkyl groupeach independently may be substituted with one or more substituentsselected from a p-toluenesulfonyloxy group, a methanesulfonyloxy group,a trifluoromethanesulfonyloxy group or a 2-tetrahydropyranyloxy,halogen, a hydroxy group and an —O-lower alkyl group-O-lower alkyl group(the alkyl group each independently may be substituted with halogen)),R^(a) and R^(b) independently represents hydrogen or a lower alkyl group(the alkyl group and the lower alkyl group each independently may besubstituted with one or more substituents selected from ap-toluenesulfonyloxy group, a methanesulfonyloxy group, atrifluoromethanesulfonyloxy group or a 2-tetrahydropyranyloxy, halogenand a hydroxy group), m is an integer of 0 to 4, and n is an integer of0 to 4, provided that at least one of R², R³ and R⁶ represents an—O-lower alkyl group (the lower alkyl group is substituted with ap-toluenesulfonyloxy group (tosyloxy group, TsO), a methanesulfonyloxygroup, a trifluoromethanesulfonyloxy group or a 2-tetrahydropyranyloxy(OTHP), and may also be substituted with one or more substituentsselected from halogen and a hydroxy group), or a pharmaceuticallyacceptable salt or solvate thereof.
 58. The compound according to claim57, wherein at least one of R², R³ and R⁶ is a group of the formula:


59. A kit for producing the labeled compound according to claim 48, or apharmaceutically acceptable salt or solvate thereof, said kitcomprising: the compound according to claim 57, or a pharmaceuticallyacceptable salt or solvate thereof, a labeling agent, and optionally,instructions for carrying out labeling.
 60. A method for producing thelabeled compound according to claim 48, which comprises the step ofreacting the compound according to claim 57, with a labeling agent.